Combined pitavastatin and dacarbazine treatment activates apoptosis and autophagy resulting in synergistic cytotoxicity in melanoma cells

Melanoma is an aggressive skin cancer and its incidence is increasing faster than any other type of cancer. Whilst dacarbazine (DTIC) is the standard chemotherapy for metastatic melanoma, it has limited success. Statins, including pitavastatin, have been demonstrated to have a range of anti-cancer effects in a number of human cancer cell lines. The present study therefore explored the anti-cancer activity of combined DTIC and pitavastatin in A375 and WM115 human melanoma cells. Cell survival assays demonstrated that combined DTIC and pitavastatin treatment resulted in synergistic cell death. Cell cycle analyses further revealed that this combined treatment resulted in a G1 cell cycle arrest, as well as a sub-G1 population, indicative of apoptosis. Activation of apoptosis was confirmed by Annexin V-fluorescein isothiocyanate/propidium iodide double-staining and an increase in the levels of active caspase 3 and cleaved poly (ADP-ribose) polymerase. Furthermore, it was demonstrated that apoptosis occurs through the intrinsic pathway, evident from the release of cytochrome c. Finally, combined DTIC and pitavastatin treatment was demonstrated to also activate autophagy as part of a cell death mechanism. The present study provides novel evidence to suggest that the combined treatment of DTIC and pitavastatin may be effective in the treatment of melanoma.Melanoma is an aggressive skin cancer and its incidence is increasing faster than any other type of cancer. Whilst dacarbazine (DTIC) is the standard chemotherapy for metastatic melanoma, it has limited success. Statins, including pitavastatin, have been demonstrated to have a range of anti-cancer effects in a number of human cancer cell lines. The present study therefore explored the anti-cancer activity of combined DTIC and pitavastatin in A375 and WM115 human melanoma cells. Cell survival assays demonstrated that combined DTIC and pitavastatin treatment resulted in synergistic cell death. Cell cycle analyses further revealed that this combined treatment resulted in a G1 cell cycle arrest, as well as a sub-G1 population, indicative of apoptosis. Activation of apoptosis was confirmed by Annexin V-fluorescein isothiocyanate/propidium iodide double-staining and an increase in the levels of active caspase 3 and cleaved poly (ADP-ribose) polymerase. Furthermore, it was demonstrated that apoptosis occurs through the intrinsic pathway, evident from the release of cytochrome c. Finally, combined DTIC and pitavastatin treatment was demonstrated to also activate autophagy as part of a cell death mechanism. The present study provides novel evidence to suggest that the combined treatment of DTIC and pitavastatin may be effective in the treatment of melanoma

TBX3 Role in Colorectal Cancer Treatment

The T-box family of developmentally important transcription factors has been shown to play a role in the genesis of cancer and shows much promise for a targeted therapeutic approach. For example, the T-box factor TBX3 is overexpressed in a number  of  cancers  including  colorectal cancer  but  its precise role  in  the  treatment of  this  disease  still need  to  be  clarified. We provide novel evidence to show that knocking down TBX3 sensitizes colorectal cancer cells to cisplatin treatment

Synergistic anticancer effect of combination treatment of vitamin D and pitavastatin on the HCC1937 breast cancer cells

Vitamin D (Vit D) has anticancer properties including activating cell senescence inhibiting cancer cell proliferation, inducing apoptotic cell death, and decreasing cancer cell migration. On the other hand, statins showed favorable anticancer activities including anti-survival, anti-proliferation, and anti-migration effects. The current study aimed to investigate the synergistic anticancer effect of Vit D and statins against HCC1937 triple-negative breast cancer cells. The antiproliferative effect was tested by MTT assay after 48 hours of the treatments. Trypan blue test and clonogenic assay were used to test the anti-survival activities of the treatments. The ability of the treatments to inhibit the migration ability was tested by scratch assay. Levels of the cell cycle and apoptotic markers were determined by western blotting. Results of the study revealed that all the tested compounds including Vit D, atorvastatin (Ator), simvastatin (Simv), and pitavastatin (Pita) inhibited HCC1937 breast cancer cell growth with different IC50 values ranging from 4.49-12.95 µM. Combined application of Pita and Vit D showed potent synergistic antiproliferative activities against HCC1937 breast cancer cells. The combined therapy of (1µM Vit D and 2 µM Pita) inhibited HCC1937 cell proliferation by cell cycle arrest and apoptosis as evidenced by increasing p21, p53, and cleaved PARP. Finally, the combined treatment decreased the p-STAT3 level in HCC1937 breast cancer cells. The results of the study can be concluded that the combined treatment of Pita and Vit D has a synergistic anticancer effect against HCC1937 breast cancer cells

Guidelines for the use and interpretation of assays for monitoring autophagy (4th edition)1.

In 2008, we published the first set of guidelines for standardizing research in autophagy. Since then, this topic has received increasing attention, and many scientists have entered the field. Our knowledge base and relevant new technologies have also been expanding. Thus, it is important to formulate on a regular basis updated guidelines for monitoring autophagy in different organisms. Despite numerous reviews, there continues to be confusion regarding acceptable methods to evaluate autophagy, especially in multicellular eukaryotes. Here, we present a set of guidelines for investigators to select and interpret methods to examine autophagy and related processes, and for reviewers to provide realistic and reasonable critiques of reports that are focused on these processes. These guidelines are not meant to be a dogmatic set of rules, because the appropriateness of any assay largely depends on the question being asked and the system being used. Moreover, no individual assay is perfect for every situation, calling for the use of multiple techniques to properly monitor autophagy in each experimental setting. Finally, several core components of the autophagy machinery have been implicated in distinct autophagic processes (canonical and noncanonical autophagy), implying that genetic approaches to block autophagy should rely on targeting two or more autophagy-related genes that ideally participate in distinct steps of the pathway. Along similar lines, because multiple proteins involved in autophagy also regulate other cellular pathways including apoptosis, not all of them can be used as a specific marker for bona fide autophagic responses. Here, we critically discuss current methods of assessing autophagy and the information they can, or cannot, provide. Our ultimate goal is to encourage intellectual and technical innovation in the field

Newly Synthesized Palladium (II) Complex ASH10 Induces Apoptosis and Autophagy in Breast Cancer Cells

Background and Objective: Palladacycles have been reported to exert significant anticancer activities against different cancer cells. The current study conducted to evaluate the anti-tumor activity of a new palladacycle complex (ASH10) in estrogen receptor-positive (MCF7) and triple-negative (HCC1937) breast cancer cells. Materials and Methods: The effect of ASH10 on cell proliferation was tested by MTT assay. Scratch assay was used to test the anti-migration ability of ASH10. Apoptosis induced by ASH10 was measured by different methods including nuclear staining and detection of apoptosis markers by western blotting. Autophagy induced by ASH10 also measured by LC3II puncta staining and western blotting using the LC3II antibody. Results: The MTT results showed that the ASH10 compound has a strong anti-growth effect. Data showed that ASH10 induces its cytotoxic effect by inducing DNA damage followed by cell cycle arrest, intrinsic apoptosis and autophagy. Compared to untreated cells, ASH10 treated cells showed high levels of DNA damage markers p-H2AX, p-ATM and p53. Importantly, the cell cycle arrest marker p21 was similarly induced by ASH10 treatment. Furthermore, ASH10 induced significant levels of apoptosis as evidenced by nuclear fragmentation and an increase in the levels of PARP cleavage. Interestingly, ASH10 also activated the formation of autophagosomes and increased the autophagy marker LC3II. Inhibition of autophagy led to a decrease in ASH10 cytotoxicity suggesting that ASH10 induced autophagy was a cytotoxic mechanism. Conclusion: These results demonstrated that ASH10 is a potential effective compound in the treatment of both ER-negative and ER-positive breast cancer cells.Background and Objective: Palladacycles have been reported to exert significant anticancer activities against different cancer cells. The current study conducted to evaluate the anti-tumor activity of a new palladacycle complex (ASH10) in estrogen receptor-positive (MCF7) and triple-negative (HCC1937) breast cancer cells. Materials and Methods: The effect of ASH10 on cell proliferation was tested by MTT assay. Scratch assay was used to test the anti-migration ability of ASH10. Apoptosis induced by ASH10 was measured by different methods including nuclear staining and detection of apoptosis markers by western blotting. Autophagy induced by ASH10 also measured by LC3II puncta staining and western blotting using the LC3II antibody. Results: The MTT results showed that the ASH10 compound has a strong anti-growth effect. Data showed that ASH10 induces its cytotoxic effect by inducing DNA damage followed by cell cycle arrest, intrinsic apoptosis and autophagy. Compared to untreated cells, ASH10 treated cells showed high levels of DNA damage markers p-H2AX, p-ATM and p53. Importantly, the cell cycle arrest marker p21 was similarly induced by ASH10 treatment. Furthermore, ASH10 induced significant levels of apoptosis as evidenced by nuclear fragmentation and an increase in the levels of PARP cleavage. Interestingly, ASH10 also activated the formation of autophagosomes and increased the autophagy marker LC3II. Inhibition of autophagy led to a decrease in ASH10 cytotoxicity suggesting that ASH10 induced autophagy was a cytotoxic mechanism. Conclusion: These results demonstrated that ASH10 is a potential effective compound in the treatment of both ER-negative and ER-positive breast cancer cells

Pitavastatin Enhances Doxorubicin-induced Apoptosis in MCF7 Breast Cancer Cells

Breast cancer is the most common malignancy in women worldwide. While doxorubicin is part of the standard therapy for metastatic breast cancer, it has limited success. Pitavastatin has been shown to enhance the anti-cancer activity of certain therapeutics. The current study, therefore, explored the anti-cancer activity of the combined treatment of doxorubicin and pitavastatin in MCF7 breast cancer cells. Cell proliferation and viability assays demonstrated that combined doxorubicin and pitavastatin treatment resulted in synergistic cytotoxicity and cell death. Western blotting analysis showed that pitavastatin treatment resulted in increasing levels of p53 and the cell cycle regulator p21 in both doxorubicin treated and untreated cells. Furthermore, we demonstrated that apoptosis induced by the combined treatment occurs through the intrinsic pathway as evident from the activation of caspase 9, caspase 7 and the reduction of BCL-2 level. This study provides novel evidence to suggest that combined treatment of doxorubicin and pitavastatin may be effectively combined to treat breast cancer with the potential to minimize the side effects associated with high doses of doxorubicin.Breast cancer is the most common malignancy in women worldwide. While doxorubicin is part of the standard therapy for metastatic breast cancer, it has limited success. Pitavastatin has been shown to enhance the anti-cancer activity of certain therapeutics. The current study, therefore, explored the anti-cancer activity of the combined treatment of doxorubicin and pitavastatin in MCF7 breast cancer cells. Cell proliferation and viability assays demonstrated that combined doxorubicin and pitavastatin treatment resulted in synergistic cytotoxicity and cell death. Western blotting analysis showed that pitavastatin treatment resulted in increasing levels of p53 and the cell cycle regulator p21 in both doxorubicin treated and untreated cells. Furthermore, we demonstrated that apoptosis induced by the combined treatment occurs through the intrinsic pathway as evident from the activation of caspase 9, caspase 7 and the reduction of BCL-2 level. This study provides novel evidence to suggest that combined treatment of doxorubicin and pitavastatin may be effectively combined to treat breast cancer with the potential to minimize the side effects associated with high doses of doxorubicin

A novel binuclear palladacycle complex inhibits melanoma growth <i>in vitro</i> and <i>in vivo</i> through apoptosis and autophagy

Please help populate SUNScholar with the full text of SU research output. Also - should you need this item urgently, please send us the details and we will try to get hold of the full text as quick possible. E-mail to [email protected]. Thank you.Journal Articles (subsidised)NatuurwetenskappeChemie & Polimeerwetenska