New Data of the Cosmeceutical and TriPeptide GHK
SOFW Journal (2015)
GHK-Cu May Prevent Oxidative Stress in Skin
by Regulating Copper and Modifying Expression of
Numerous Antioxidant Genes Cosmetics (2015)
GHK Peptide as a Natural Modulator of
Multiple Cellular Pathways in Skin Regeneration (2015)
GHK, the Human Skin Remodeling Peptide Induces Anti-Cancer
Expression and DNA Repair Analytical Oncology (2014)
GHK and DNA: Resetting the Human Genome to Health
BioMed Research International (2014)
Avoid Buying Fake Copper Peptides Dangerous



GHK Suppresses Cancer Metastasis Genes, Increases Cancer Suppressors Decorin and Protein P63, and Programmed Cell Death (Apoptosis) of Cancer Cells, and Growth Inhibitory Analogs of the GHK Copper Binding Region

This information is for the use of medical researchers and persons interested in health sciences. It must be emphasized that GHK and other copper binding peptides have not been tested on human cancers and are not approved for such uses.

GHK's anti-cancer effect seems to be tied to its effects on cell differetiation. Cancer cells tend to be a poorly differentiatedand more like stem cells. Peled et al found GHK with added copper to strongly caused diffentiation of hematopoietic stem cells whereas GHK without copper did not cause differentiation. (Methods of controlling proliferation and differentiation of stem and progenitor cells, United States Patent: 6,962,698, Peled, Tony, Fibach, Eitan, Treves; Avi, Gamida Cell Ltd. (Jerusalem, IL) and Hadasit Medical Research Services and Development, Ltd. (Jerusalem, IL))

Our work indicated that "copper-free" growth medium still contained about 50 to 300 nanomolar copper as a trace contaminant. So GHK in a low copper system does not cause differentiation, but GHK with signficant added copper causes differentiation.

The GHK-copper 2+ effects are similar to retinoic acid. These two compounds are in the human body and possess both skin regenerative actions and strong anti-cancer actions.

How Cancer Research Went Down the Wrong Path

Progress in cancer research over the past 50 years has pathetic. Today's most promising anti-cancer drugs may prolong your life a few months, cost up to $1,000 a day, and cause you great misery. 50 years ago there were debates on the best approach to use to treat cancer. There was the sledgehammer approach of poisons and radiation treatments used to kill the cells. On the other side were a few scientists who argued that be best anti-cancer drugs were actually causing celluar differentiation of the cancer cells to a stage where they stopped uncontrolled growth. Non-scientists tend to think of science research as a rational field but in reality it is more like a buffalo stampede where a movement starts and everyone goes out-of-control in the same direction. If you doubt me, think of the advice over the past 50 years on cholesterol where all those years we were taught the dangers of cholesterol, but now it is not considered a cause of cardiovascular disease.

GHK and retinoic acid may open a door to controlling cancers with natural molcules from the human body.

It is very possible that GHK could be used to develop highly effective and non-toxic therapies for human cancer. There is very little interest in non-toxic cancer therapies because of the widespread belief in the cancer treatment field that cancers must be destroyed with extremely damaging poisons and radiation.

Cancer Suppression with Vitamin C and Copper:Gly-Gly-His

This 1983 paper by Linus Pauling's group found that a copper peptide very similar to GHK gave a strong tumor inhibition when administered with Vitamin C. The non-toxic procedure either extended the lifespan of tumor-bearing mice or killed the tumors. All of the untreated tumor-bearing mice died. 60% of the treated mice lived twice as long as the tumor-only mice and 40% appear to have been cured of the cancer. The tumor could not be re-implanted in the surviving mice and the paper said "..the mice lived a for a long period."

GGH-Cu and Vitamin C Suppress Cancer

Cancer Research. 1983 Feb;43(2):824-28.

Enhancement of Antitumor Activity of Ascorbate against Ehrlich Ascites Tumor Cells by the Copper:Glycylglycylhistidine Complex

Kimoto E, Tanaka H, Gyotoku J, Morishige F, Pauling L, Department of Chemistry, Faculty of Science, Fukuoka University, Fukuoka 814, Japan [E. K., H. T., J. G.]; Tachiarai Hospital, Fukuoka 838, Japan IF. M.]," and Linus Pauling Institute of Science and Medicine, Palo Alto, California 94306 [L. P.]

Abstract

Ascorbate in an aqueous solution is easily oxidized by molecular oxygen in the presence of cupric ion, thus producing reactive oxygen species and exhibiting cytotoxicity.

In order to increase the antitumor activity of ascorbate, we used the innocuous form of cupric ion complexed with glycyl- glycylhistidine, a tripeptide designed to mimic the specific Cu(ll) transport site of albumin molecule. Although this square planar copper:glycylglycylhistidine complex did not significantly oxidize ascorbate at pH 7.4, it killed Ehrlich ascites tumor cells in vitro in a high concentration of ascorbate. The injections of large doses of ascorbate together with copper: glycylglycylhistidine prolonged the life span of mice inoculated i.p. with Ehrlich tumor cells. The target specificity against tumor cells was primarily attributable to their high peptide-cleaving activity.

 

 

Repeat of Pauling Experiment with GHK-Cu and Vitamin C in mice with fibrosarcoma

In 1986, I repeated the Pauling anti-cancer treatment in mice but with GHK-copper and vitamin C. The mice were implanted with fibrosarcoma cells which grows as a solid mass in the thigh muscles. After 5 days, the mice were injected intraperitoneally with GHK-copper and Vitamin C with the Pauling dosage three times per week (Mon, Wed, Fri) for 2 weeks. Then the experiment was stopped at 6 weeks.

There was a strong inhibition of tumor growth (about -70%) and some of the mice appeared
to have no tumors. The injected copper peptides and vitamin C appeared to have no negative
effect on the mice's behavior or physical condition. The first row is the control mice. The section is the mice with tumors which show as an increase in the muscle tissue. The third row is mice with the tumors which were treated with vitamin C and GHK-copper.

I could not obtain funding to extend this work from the National Cancer Institute. The Review Panel said such therapies were not needed since new non-toxic biologicals would be used in the near future to cure cancer. However, 26 years later, damaging and poorly effective toxic cancer therapies are still the main treatment.


DrPickarts NEW 2014 Paper on GHK and Cancer Suppression
The Effect of GHK-Cu and Ascorbic Acid on the Growth of Scarcoma in Mice


GHK Suppression of Cancer Metastasis Genes

2010, Hong et al, using the the Broad Institute's Connectivity Map, predicted that GHK, at a low and non-toxic level, would reverse many of the effects of aggressive, metastatic human colon cancer on 40 critical cancer-spreading genes.

The gene test used GHK without copper but it is likely that the much of the GHK was converted to GHK-copper because of the extremely high affinity of GHK for copper. Part of this effect may have been due to GHK-copper's effect on increasing the protein Decorin (Named because it "decorates" the collagen strands). Decorin has a perplexing mixture of effects: Regeneration of muscles and nerves, reduction of scarring, and inhibition of tumor growth and cancer metastasis in animals.

The authors of this study searched for substances that can reverse expression of those genes involved in human colon cancer metastasis.

To determine gene expression of human normal and cancerous cells, they measured RNA produced by cells. To find substances that can reverse expression of those genes involved in metastasis, they used very reliable database developed at the Broad Institute (Massachusetts Institute of Technology and Harvard) of 7000 genome wide expression profiles after treatment of 4 human cell lines with 1309 bioactive substances. Only two substances, GHK at 1 micromolar and securenine (an alkaloid) at 18 micromolar from the choice of 1309 were able to quiet down the genes involved in tumor spreading. The authors also mention low toxicity of GHK and the low concentration that produces desired effect.

To determine GHK's potential as a cancer treatment, we analyzed the molecule's effect on the human gene expression. We found that GHK induces anti-cancer expression of numerous caspase, growth regulatory, and DNA repair genes. Below is some of our data:

GHK and Gene Expression in Apoptosis Proteins

Genes

Percent Change in Gene Expression

Comment

CASP 1

432

Caspase proteins activate programmed cell death

CASP 3*

65


CASP 6*

23


CASP 7*

48


CASP 8

399


CASP 10

195


NLRP1

249

Apoptosis caspase recruitment domain

CARD10

173

Apoptosis signaling gene

BCL2L14

153

Apoptosis facilitator


GHK and Gene Expression in Cancer Suppressors

Genes

Percent Change in Gene Expression

Comment

USP29

1056

Ubiquitin specific peptidase 29,

May stabilize P53 tumor suppressor

IFNA21

955

Combined treatment of IFN-alpha and IL-21 increases anti-cancer effects

TP73

938

Tumor suppressor

TP63

Uncertain

Tumor suppressor

Gene probes are inconsistent however, TP63 was induced by GHK in keratinocyte cells in skin equivalent organ culture

LEFTY2

935

Inhibition of pancreatic cancer cells

IL25

891

Inhibits breast cancer cell growth

IL15

875

Induces natural killer cells

Anti-tumor and anti-viral

D4S234E


731

p53-responsive gene, induces apoptosis in response to DNA damage

MTUS2

474

Microtubule associated tumor suppressor

C13orf18

352

Inhibits cervical cancer cells

ING2

337

Functions in DNA repair and apoptosis

CTNNA1

336

Suppresses cancer invasion of tissues

CDKN1C

277

Breast cancer inhibitor

PAWR

199

Induces apoptosis in cancer cells

APC

195

Suppresses colon cancer

PTEN

165

Cancer suppressor

NRG1

164

Cancer suppressor

NF1

143

Neurofibromin 1

ATM

107

Senses DNA damage

ING4

107

Cancer suppressor

DCN*

44

Suppresses cancer growth and metastasis

In rat wound chamber experiments GHK increased mRNA for decorin 302%

BRCA1*

44

Cancer suppressor


GHK and Gene Expression in Cancer Enhancers

Genes

Percent Change in Gene Expression

Comment

ABCB1

-1537

Increases drug resistance in cancer cells

STAT5

-982

Signals cancer cells to grow

FGFR2

-904

FGFR2 inhibitors reduce some cancers

FAIM2

-749

Prevents apoptosis

IGF1

-522

Risk factor for cancer

TNF

-115

May promote cell cancer invasion

 

 

GHK suppresses human cancer metastasis genes

Clin Exp Metastasis. 2010 Feb;27(2):83-90. Epub 2010 Feb 9.

A 'metastasis-prone' signature for early-stage mismatch-repair proficient sporadic colorectal cancer patients and its implications for possible therapeutics.

Hong Y, Downey T, Eu KW, Koh PK, Cheah PY.,Department of Colorectal Surgery, Singapore General Hospital, Singapore, 169608, Singapore.

Abstract

Metastasis is the major cause of cancer mortality. We aimed to find a metastasis-prone signature for early stage mismatch-repair proficient sporadic colorectal cancer (CRC) patients for better prognosis and informed use of adjuvant chemotherapy. The genome-wide expression profiles of 82 age-, ethnicity- and tissue-matched patients and healthy controls were analyzed using the Affymetrix U133 Plus 2 array. Metastasis-negative patients have 5 years or more of follow-up. A 10 x 10 two-level nested cross-validation design was used with several families of classification models to identify the optimal predictor for metastasis. The best classification model yielded a 54 gene-set (74 probe sets) with an estimated prediction accuracy of 71%. The specificity, sensitivity, negative and positive predictive values of the signature are 0.88, 0.58, 0.84 and 0.65, respectively, indicating that the gene-set can improve prognosis for early stage sporadic CRC patients. These 54 genes, including node molecules YWHAB, MAP3K5, LMNA, APP, GNAQ, F3, NFATC2, and TGM2, integrate multiple bio-functions in various compartments into an intricate molecular network, suggesting that cell-wide perturbations are involved in metastasis transformation. Further, querying the; 'Connectivity Map' with a subset (70%) of these genes shows that Gly-His-Lys and securinine could reverse the differential expressions of these genes significantly, suggesting that they have combinatorial therapeutic effect on the metastasis-prone patients. These two perturbagens promote wound-healing, extracellular matrix remodeling and macrophage activation thus highlighting the importance of these pathways in metastasis suppression for early-stage CRC.

Wound Healing Society Presentation

Wound Healing Society Presentation

Click here for full poster

 

GHK-copper increases decorin which suppresses cancer growth and metastasis

GHK-copper increases Decorin J Invest Dermatol. 2000 Dec;115(6):962-8.
Expression of glycosaminoglycans and small proteoglycans in wounds: modulation by the tripeptide-copper complex glycyl-L-histidyl-L-lysine-Cu(2+).
Siméon A, Wegrowski Y, Bontemps Y, Maquart FX. Laboratoire de Biochimie Médicale et de Biologie Moléculaire, FRE CNRS 2260, IFR 53-Biomolécules, Faculté de Médecine, Reims, France.

Glycyl-histidyl-lysine-Cu(2+) is a tripeptide-copper complex previously shown to be an activator of wound healing. We have investigated the effects of glycyl-histidyl-lysine-Cu(2+) on the synthesis of glycosaminoglycans and small proteoglycans in a model of rat experimental wounds and in rat dermal fibroblast cultures. Repeated injections of glycyl-histidyl-lysine-Cu(2+) (2 mg per injection) stimulated the wound tissue production, as appreciated by dry weight and total protein measurements. This stimulation was accompanied by an increased production of type I collagen and glycosaminoglycans (assessed, respectively, by hydroxyproline and uronic acid contents of the chamber). Electrophoretic analysis of wound tissue glycosaminoglycans showed an accumulation of chondroitin sulfate and dermatan sulfate in control wound chambers, whereas the proportion of hyaluronic acid decreased with time. The accumulation of chondroitin sulfate and dermatan sulfate was enhanced by glycyl-histidyl-lysine-Cu(2+) treatment. The expression of two small proteoglycans of the dermis, decorin and biglycan, was analyzed by northern blot. The biglycan mRNA steady-state level in the chamber was maximal at day 12, whereas the decorin mRNA increased progressively until the end of the experiment (day 22). Glycyl-histidyl-lysine-Cu(2+) treatment increased the mRNA level of decorin and decreased those of biglycan. In dermal fibroblast cultures, the stimulation of decorin expression by glycyl-histidyl-lysine-Cu(2+) was also found. In contrast, biglycan expression was not modified. These results show that the expression of different proteoglycans in wound tissue are regulated in a different manner during wound healing. The glycyl-histidyl-lysine-Cu(2+) complex is able to modulate the expression of the extracellular matrix macromolecules differently during the wound repair process.
Decorin suppresses growth and metastasis of cancer cells Sheng Wu Yi Xue Gong Cheng Xue Za Zhi. 2007 Feb;24(1):222-5.
[The action of decorin in anti-fibrosis and anti-cancer]
[Article in Chinese]
Ma W, Tan Y, Cai S, Chen H, Du J, Cai S.
Key Laboratory for Biomechanics & Tissue Engineering of the State Ministry of Education, College of Bioengineering, Chongqing University, Chongqing 400044, China.

Decorin (DCN) is a member of the small leucine-rich proteoglycan gene family. Many studies indicated that DCN inhibited fibrosis and scar-formation by neutralization of TGF-P and interfering the binding of TGF-beta with its receptor, which induced ectopic deposition of extracellular matrix. Additionally, DCN can prevent the proliferation and metastasis of tumor cells by activating EGFR/MAPK/p21 signal pathway and inhibiting the cell proliferation pathway mediated by EGF-EGFR. It is suggested that the recombinant DCN had potential pharmaceutical potency in treatment of chronic fibrosis and neoplasm for its critical biological activities and low immunogenicity.
Decorin prevents metastatic spreading of breast cancer Oncogene. 2005 Feb 3;24(6):1104-10.
Decorin prevents metastatic spreading of breast cancer.
Reed CC, Waterhouse A, Kirby S, Kay P, Owens RT, McQuillan DJ, Iozzo RV.
Department of Pathology, Anatomy and Cell Biology, Thomas Jefferson University, Philadelphia, PA 19107, USA.

Metastases in breast cancer are a vital concern in treatment, with epidermal growth factor receptor and ErbB2 strongly implicated in mediating tumor invasion and spreading. In this study, we investigated the role of decorin in suppressing both primary breast carcinomas and pulmonary metastases. We show that decorin causes marked growth suppression both in vitro and in vivo using a metastatic breast cancer cell line and an orthotopic mammary carcinoma model. Treatment with decorin protein core reduced primary tumor growth by 70% and eliminated observed metastases. An adenoviral vector containing the decorin transgene caused primary tumor retardation of 70%, in addition to greatly reducing observed metastases. Moreover, we demonstrate that ErbB2 phosphorylation and total receptor protein levels are reduced in this model system upon de novo expression of decorin under the control of a doxycycline-inducible promoter. Primary tumor growth in vivo was reduced by up to 67% upon decorin induction, and pulmonary metastases were markedly hampered as well. These effects are likely occurring through decorin's long-term downregulation of the ErbB2 tyrosine kinase cascade. These results demonstrate a novel role for decorin in reduction or prevention of tumor metastases in this breast cancer model and could eventually lead to improved therapeutics for metastatic breast cancer.

Decorin prevents metastatic spreading of breast cancer Am J Pathol. 2008 Sep;173(3):844-55. Epub 2008 Aug 7.
An antimetastatic role for decorin in breast cancer.
Goldoni S, Seidler DG, Heath J, Fassan M, Baffa R, Thakur ML, Owens RT, McQuillan DJ, Iozzo RV.
Department of Pathology, Anatomy and Cell Biology, Thomas Jefferson University, Philadelphia, PA 19107, USA.

Decorin, a member of the small leucine-rich proteoglycan gene family, down-regulates members of the ErbB receptor tyrosine kinase family and attenuates their signaling, leading to growth inhibition. We investigated the effects of decorin on the growth of ErbB2-overexpressing mammary carcinoma cells in comparison with AG879, an established ErbB2 kinase inhibitor. Cell proliferation and anchorage-independent growth assays showed that decorin was a potent inhibitor of breast cancer cell growth and a pro-apoptotic agent. When decorin and AG879 were used in combination, the inhibitory effect was synergistic in proliferation assays but only additive in both colony formation and apoptosis assays. Active recombinant human decorin protein core, AG879, or a combination of both was administered systemically to mice bearing orthotopic mammary carcinoma xenografts. Primary tumor growth and metabolism were reduced by approximately 50% by both decorin and AG879. However, no synergism was observed in vivo. Decorin specifically targeted the tumor cells and caused a significant reduction of ErbB2 levels in the tumor xenografts. Most importantly, systemic delivery of decorin prevented metastatic spreading to the lungs, as detected by novel species-specific DNA detection and quantitative assays. In contrast, AG879 failed to have any effect. Our data support a role for decorin as a powerful and effective therapeutic agent against breast cancer due to its inhibition of both primary tumor growth and metastatic spreading.

Decorin suppresses lung cancer metastasis Oncol Rep. 2008 Jun;19(6):1533-9.
Decorin suppresses lung metastases of murine osteosarcoma.
Shintani K, Matsumine A, Kusuzaki K, Morikawa J, Matsubara T, Wakabayashi T, Araki K, Satonaka H, Wakabayashi H, Iino T, Uchida A.
Department of Orthopaedic Surgery, Mie University Faculty of Medicine, Mie, Japan.

Lung metastasis is the most crucial event affecting the therapeutic outcome of osteosarcoma. The prevention of lung metastasis is therefore important in improving the prognosis of patients with osteosarcoma. Decorin is a major extracellular matrix protein which has become the focus of various cancer studies. The biological role of decorin in osteosarcoma has yet to be clarified. The aim of this study was to examine the potential of decorin as a novel biological target for the treatment of osteosarcoma. In this study, the LM8 murine osteosarcoma cell line (LM8) with high metastatic potential to the lung was used. The two cell lines established were LM8-DCN which stably expressed human decorin (hDCN) and LM8-mock, established as a control. The LM8-DCN cell line was subcutaneously injected into the backs of mice. Significantly fewer pulmonary metastases were observed in mice with LM8-DCN compared to mice inoculated with LM8 and LM8-mock (P<0.001). In addition, the mice in the LM8-DCN inoculated group survived significantly longer than those in the LM8 and LM8-mock inoculated group, based on the Kaplan-Meier survival analysis and log-rank tests (P<0.005). The effect of decorin on the growth rates, motility and invasion ability of LM8 was investigated in vitro. There was no difference in the morphology and growth rates, but the motility and invasion of LM8 were inhibited by decorin. These results suggest that decorin has the therapeutic potential to prevent lung metastasis in osteosarcoma.

Decorin suppesses breast cancer metastasis into bone

Oncology. 2009;77(2):92-9. Epub 2009 Jul 7.
Decorin suppresses bone metastasis in a breast cancer cell line.

Araki K, Wakabayashi H, Shintani K, Morikawa J, Matsumine A, Kusuzaki K, Sudo A, Uchida A. Department of Orthopaedic Surgery, Mie University Graduate School of Medicine, Mie, Japan.

Decorin, the prototype of an expanding family of small leucine-rich proteoglycans, is involved in a number of cellular processes including matrix assembly, fibrillogenesis and the control of cell proliferation. In this study, we investigated the role of decorin in suppressing tumor aggressiveness and bone metastases. We used a metastatic breast cancer cell line, MDA-MB-231, to show that decorin causes marked growth suppression bothin vitro and in vivo. A cytomegaloviral vector containing the decorin transgene caused greatly reduced cell growth, motility and observed metastases. Bone metastases were decreased by >90% upon decorin transfection. These results demonstrate a novel role for decorin in the reduction or prevention of tumor metastases in this breast cancer model and could eventually lead to improved therapies for metastatic breast cancer.

Decorin suppesses prostate cancer growth Neoplasia. 2009 Oct;11(10):1042-53.
Decorin suppresses prostate tumor growth through inhibition of epidermal growth factor and androgen receptor pathways.
Hu Y, Sun H, Owens RT, Wu J, Chen YQ, Berquin IM, Perry D, O'Flaherty JT, Edwards IJ.
Department of Pathology, Wake Forest University School of Medicine, Winston-Salem, NC 27157, USA.

Epidermal growth factor receptor (EGFR) and androgen receptor (AR) pathways play pivotal roles in prostate cancer progression. Therefore, agents with dual-targeting ability may have important therapeutic potential. Decorin, a proteoglycan present in the tumor microenvironment, is known to regulate matrix assembly, growth factor binding, and receptor tyrosine kinase activity. Here, we show that in prostate-specific Pten(P-/-) mice, a genetically defined, immune-competent mouse model of prostate cancer, systemic delivery of decorin inhibits tumor progression by targeting cell proliferation and survival pathways. Moreover, in human prostate cancer cells, we show that decorin specifically inhibits EGFR and AR phosphorylation and cross talk between these pathways. This prevents AR nuclear translocation and inhibits the production of prostate specific antigen. Further, the phosphatidylinositol-3 kinase (PI3K)/Akt cell survival pathway is suppressed leading to tumor cell apoptosis. Those findings highlight the effectiveness of decorin in the presence of a powerful genetic cancer risk and implicate decorin as a potential new agent for prostate cancer therapy by targeting EGFR/AR-PI3K-Akt pathways.

Decorin inhibits primary tumor growth and metastatic speading Int J Cancer. 2008 Dec 1;123(11):2473-9.
Tumor microenvironment: Modulation by decorin and related molecules harboring leucine-rich tandem motifs.
Goldoni S, Iozzo RV.
Department of Pathology, Anatomy and Cell Biology, and the Cancer Cell Biology and Signaling Program, Kimmel Cancer Center, Thomas Jefferson University, Philadelphia, PA 19107, USA.

Decorin, the prototype member of the small leucine-rich proteoglycans, resides in the tumor microenvironment and affects the biology of various types of cancer by downregulating the activity of several receptors involved in cell growth and survival. Decorin binds to and modulates the signaling of the epidermal growth factor receptor and other members of the ErbB family of receptor tyrosine kinases. It exerts its antitumor activity by a dual mechanism: via inhibition of these key receptors through their physical downregulation coupled with attenuation of their signaling, and by binding to and sequestering TGFbeta. Decorin also modulates the insulin-like growth factor receptor and the low-density lipoprotein receptor-related protein 1, which indirectly affects the TGFbeta receptor pathway. When expressed in tumor xenograft-bearing mice or injected systemically, decorin inhibits both primary tumor growth and metastatic spreading. In this review, we summarize the latest reports on decorin and related molecules that are relevant to cancer and bring forward the idea of decorin as an anticancer therapeutic and possible prognostic marker for patients affected by various types of tumors. We also discuss the role of lumican and LRIG1, a novel cell growth inhibitor homologous to decorin.

P63 as a Cancer Inhibitor

There is some evidence that protein P63 also inhibits cancer growth. GHK-copper increases protein P63.

GHK-copper increases protein P63

Arch Dermatol Res. 2009 Apr;301(4):301-6. Copper-GHK increases integrin expression and p63 positivity by keratinocytes. Kang YA, Choi HR, Na JI, Huh CH, Kim MJ, Youn SW, Kim KH, Park KC. Department of Dermatology, Seoul National University College of Medicine, Yeongeon-dong, Jongno-gu, Seoul, Republic of Korea.

Glycyl-L-histidyl-L-lysyl (GHK) possesses a high affinity for copper(II) ions, with which it spontaneously forms a complex (copper-GHK). It is well known that copper-GHK plays a physiological role in the process of wound healing and tissue repair by stimulating collagen synthesis in fibroblasts. This study was conducted to investigate the effects of copper-GHK on keratinocytes. Proliferative effects were analyzed and hematoxylin and eosin staining and immunohistochemistry were conducted to evaluate the effects of copper-GHK in skin equivalent (SE) models. In addition, western blotting was performed. In monolayer cultured keratinocytes, copper-GHK increased the proliferation of keratinocytes. When the SE models were evaluated, basal cells became cuboidal when copper-GHK was added. Immunohistochemical analysis revealed that copper-GHK increased proliferating cell nuclear antigen (PCNA) and p63 positivity. Furthermore, the expression of integrin alpha6 and beta1 increased in SE models, and these results were confirmed by Western blotting. The results of this study indicate that treatment with copper-GHK may increase the proliferative potential of basal keratinocytes by modulating the expression of integrins, p63 and PCNA. In addition, increased levels of p63, a putative stem cell marker of the skin, suggests that copper-GHK promotes the survival of basal stem cells in the skin.

P63 may suppress cancer

Cell Death Differ. 2010 Apr 9. [Epub ahead of print]

p53-family proteins and their regulators: hubs and spokes in tumor suppression.
Collavin L, Lunardi A, Del Sal G. Laboratorio Nazionale CIB (LNCIB), AREA Science Park, Trieste, Italy, Dipartimento di Scienze della Vita, Università degli Studi di Trieste, Trieste, Italy.

Abstract
The tumor suppressor p53 is a central hub in a molecular network controlling cell proliferation and death in response to potentially oncogenic conditions, and a wide array of covalent modifications and protein interactions modulate the nuclear and cytoplasmic activities of p53. The p53 relatives, p73 and p63, are entangled in the same regulatory network, being subject at least in part to the same modifications and interactions that convey signals on p53, and actively contributing to the resulting cellular output. The emerging picture is that of an interconnected pathway, in which all p53-family proteins are involved in the response to oncogenic stress and physiological inputs. Therefore, common and specific interactors of p53-family proteins can have a wide effect on function and dysfunction of this pathway. Many years of research have uncovered an impressive number of p53-interacting proteins, but much less is known about protein interactions of p63 and p73. Yet, many interactors may be shared by multiple p53-family proteins, with similar or different effects. In this study we review shared interactors of p53-family proteins with the aim to encourage research into this field; this knowledge promises to unveil regulatory elements that could be targeted by a new generation of molecules, and allow more efficient use of currently available drugs for cancer treatment.

   

GHK Induces Programmed Cell Death in Cancer Cells

Cancer cells survive by shutting down the systems that kill cancer cells using programmed cell death or apoptosis. The paper by Hong et al also reported that the cancer cells were able to shut down the cancer-killing apoptosis mechanism.

Now a 2012 paper reports that GHK, at a very low concentration, does not harm normal fibroblasts (a major repair cell) but did induce programmed cell death two cancer cell lines. Thus, GHK reversed this effect of the cancer cells on the genes.

The Tripeptide, GHK, Induces Programmed Cell Death in SH-SY5Y Neuroblastoma Cells Luay E. Matalka, Ashley Ford and M. Tino Unlap Matalka LE, Ford A, Unlap MT (2012) J Biotechnol Biomater 2:144

Abstract

GHK-Cu is a tripeptide that is found in plasma and increases fibroblast proliferation and is widely used in wound healing creams. Studies show that GHK-Cu stimulates the level of p63, a member of the antitumor suppressor family which includes p53 and p73. Because of its effect on p63, we tested the hypothesis that GHK induces apoptosis in neuroblastoma cells, SH-SY5Y, and that modification of GHK by polyethylene glycol (PEG) conjugation, PEGylation, potentiates its effects. Initial studies on NIH-3T3 fibroblasts and U937 histiocytic lymphoma cells show that GHK-Cu and GHK-PEG stimulate fibroblast proliferation while attenuating U937 cell proliferation. SH-SY5Y cells were treated with GHK or GHK-PEG at 1 and 10 nM for 24 hours followed by cell proliferation, cell viability, cell cytotoxicity and apoptosis assays. Our results showed that 24 hour GHK treatment elevated apoptosis by 1.8 ± 0.17 and 3.3 ± 0.15 fold at 1 and 10 nM, respectively. GHK-PEG treatment, on the other hand, stimulated apoptosis by 3.2 ± 0.80 and 4.9 ± 0.9 fold at 1 and 10 nM, respectively. The treatments, while having no effects on cell cytotoxicity, reduced neuroblastoma cell viability and cell proliferation. Therefore, GHK induces apoptosis in neuroblastoma cells, an affect which was potentiated by PEGylation.

The complete paper The Tripeptide, GHK, Induces Programmed Cell Death in SH-SY5Y Neuroblastoma Cells by Luay  E.  Matalka,  Ashley  Ford  and  M.  Tino  Unlap can be downloaded for free here.

Dr. Unlap has communicated to me that GHK similarily induced programmed cell death in human breast cancer cells.

The above paper measured production caspase 3 and caspase 7 as an indicator of apoptosis activity. Our results from GenePattern indicate that GHK induces 7 caspases. There are 12 known caspase proteins in humans.

Caspase Number - Fold Increase - mRNA

Caspase 1 - 5.32

Caspase 2 - 0.72 (net average of +2.89 and -2.17)

Caspase 3 - 1.65

Caspase 6 - 1.23 (net average of +1.08 and +1.38)

Caspase 7 - 1.48

Caspase 8 - 4.99

Caspase 10 - 2.95 (net average of +3.35 and +2.54)

The conflicting results for caspase 2 indicate a problem with the gene probe set. The probe sets measure only a small part of the m-RNA and are often refined to give more accurate data.


Dr. Pickart's NEW! 2014 Paper:
GHK, the Human Skin Remodeling Peptide, Induces Anti-Cancer Expression of Numerous Caspase, Growth Regulatory, and DNA Repair Genes can be downloaded for FREE - Click Here

 

I and colleagues at the University of Washington also synthesized many copper complexes similar to GHK-copper and these suppressed cancer growth in cell culture and in animals.

Anti-cancer analogs of GHK copper binding site

Inhibition of the growth of cultured cells and an implanted fibrosarcoma by aroylhydrazone analogs of the Gly-His-Lys-Cu(II) complex. Biochem Pharmacol 1983 Dec 15;32(24):3868-71.

Pickart L, Goodwin WH, Burgua W, Murphy TB, Johnson DK, Biochem. Pharmacol., 32, 1983, 3868-71.

Anti-cancer analogs of GHK copper binding site Cytotoxic chelators and chelates 1. Inhibition of DNA synthesis in cultured rodent and human cells by aroylhydrazones and by a copper(II) complex of salicylaldehyde benzoyl hydrazone. Inorganica Chimica Acta, 67:159-165, 1982.
David K. Johnson, Terrance B. Murphy, Norman J. Rose, William H. Goodwin and Loren Pickart, Department of Medicinal Chemistry and Chemistry, University of Washington, Seattle, Wash. 98195,Virginia Mason Research Center, Seattle, Wash. 98101

Abstract

Aroylhydrazones of pyridoxal and of salicylaldehyde, a series of tridentate chelating agents, are potent inhibitors of DNA synthesis and cell growth in a number of human and rodent cell lines grown in culture. A copper(II) complex of the most potent of the chelators, salicylaldehyde benzoyl hydrazone (SBH), exhibits significantly greater inhibitory activity than does SBH itself. Although the bioactive forms and mechanism of action of these agents are uncertain, their cytotoxic activity can equal or exceed that of many chelators and chelates previously known to possess such properties, including compounds used clinically. SBH and its copper complex are relatively non-toxic to mice and show some measure of selectivity in their effects on different cell types. It is possible that aroylhydrozones of this type and/or their metal complexes could prove to be useful therapeutic agents.

Anti-cancer analogs of GHK copper binding site Biological activity of human plasma copper-binding growth factor glycyl-l-histidyl-l-lysine, Pickart L, Lovejoy S, Methods Enzymol. 147, 1987, 314-328

Hydrazone analogs of the GHK copper binding region are very potent inhibitors of DNA synthesis and cell growth. Furthermore, the wound repairing cell, the fibroblast, seems to be exceptionally sensitive to certain types of copper-complex inhibitors. This makes the preparation of tissue regenerative copper complexes difficult since even some types of peptide copper complexes inhibit fibroblasts as do other small complexes such as diethylamine-copper and triethylamine-copper.GHK Inhinitors
 
 
 
 
 
 
 GHK-Cu and hydrazone analogs of GHK copper binding region that that are very potent inhibitors of cell replication

Top: Copper binding region of GHK

Middle: PCPH-Cu (2-Pyridinecarboxaldehyde 2''-pyridylhydrazone hydrazone copper(2+))

Bottom: SBH-Cu (Salicylaldehyde benzoyl hydrazone copper(2+))
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 

 

 

Cell Types in Culture

PCPH-Cu
Nanograms per ml for 50% cell reduction

SBH-Cu
Nanograms per ml for 50% cell reduction

Cisplatin
Nanograms per ml
for 50% cell reduction

Bleomycin
Nanograms per ml
for 50% cell reduction

Normal human fibroblasts

0.00003

0.1

Not done

-

Normal human kidney

45

 

-

-

Mouse fibrosarcoma

0.511

2.4

32

Human bladder cancer

12

34

95

7

Human lung epithelial cancer

580

410

870

44

Human melanoma

0.05

270

6,000

80,000

Questions or Advice?

Email Dr. Loren Pickart at drlorenpickart@gmail.com

Alternate Email: ghkcopperpeptides@gmail.com