Study on the Anti-cancer Potential of Aegle marmelos Fruit Extract Pro and Anti Apoptotic Molecule in Human Melanoma Cell Line-A375

Aegle marmelos is also known as bael which is commonly found in south East Asia and Indian-sub continent. The origin of bael is India. Bael is also known as the golden apple, Bengal-quince in India. In the ancient medical system, Aegle marmelos play an important role and its extract is also useful in inflammation, diabetes, cancer and asthma. The leaves are used for anti-inflammatory, nervous disorder, control blood sugar and fruit is used to treat antiviral, anti-diabetics, and brain and heart tonic. In addition, studies have proved that bael is used for the treatment and prevention of cancer. The main aim of this study is to assess the anti-cancer potential of Aegle marmelos fruit extract pro and anti apoptotic molecules in human melanoma cell line-A375. In the present study, Human Melanoma A375 cells will be produced, grown and will be passed in different culture flasks, then RNA isolation was done and by reverse transcriptase process, RNA gets converted into cDNA. This cDNA will be used for the amplification of growth factor beta using gene specific primers by commercially available real time PCR kit. The anticancer potential effect was found in 400µg/ml, as the concentration increases, the cell viability is decreased. The current study explains the potential application of bael in pharmacological and medicinal uses in near future.

A Review of Progesterone Effects on Human Melanoma Cell Growth In-Vitro

Progesterone, a female sex hormone not only has a role in reproduction, but also in protecting females in melanoma. A survey of steroid hormones actions steroid hormones actions survey on melanoma cells and literature survey showed that progesterone inhibited mouse and human melanoma cell growth significantly in-vitro. Progesterone not only inhibited cell growth, but also affected adhesion and migration functions (essential for metastasis) in-vitro. This observation correlated with the clinical studies where they had shown showed an increased survival and delayed metastasis in menstruating females in melanoma. Further, progesterone level in menstruating females (1000–1500 ng/dL) compared to post-menopausal females (20–100 ng/dL) also correlated with previous clinical studies. Progesterone action on melanoma cells, as reported by other researchers also supported the findings from this lab. Hence, progesterone could be the steroid hormone protecting menstruating females in melanoma. Moreover, our recent studies showed that progesterone suppressed pro-inflammatory cytokine IL-8 secretion by the melanoma cells, which decreased melanoma cell growth in-vitro. Hence, progesterone apart from reproductive function may also be involved in protecting menstruating females in melanoma.

Effect of Aegle Marmelos Hydroethanolic Leaf Extract on Expression of Antiapoptotic Markers in Human Melanoma Cells

Background: Aegle marmelos commonly known as Bael is a herbal plant. Itis from a family called Rutaceae. It has many medicinal uses: anti-diarrheal, anti- microbial, anti-viral, anti-cancer, chemo-preventive, Ulcer healing and many others. Aim: To study the effect of Aegle marmelos hydroethanolic leaf extract on expression of antiapoptotic markers in human melanoma cells. Objective: The present study investigated the effect of Aegle marmelos hydroethanolic leaf extract on expression of antiapoptotic markers in human melanoma cells. Materials and Methods: DMSO and MTT chemicals were purchased from Sigma chemical Pvt Ltd. Trypsin EDTA, FBS, RPMI 1640 medium and PBS, Real time PCR kit was purchased from Canada. Human melanoma cell line (A375) was purchased from NCCS, Pune, India. Results: The data was analysed statistically by ANOVA and Duncan’s multiple range test with a computer based software (Graph Pad Prism version 5). The percentage of cell viability decreases with the increase in dosage of Aegle marmelos leaf extract. Conclusion: The study concluded that Aegle marmelos hydro ethanolic leaf extract a novel and innovative herbal drug has a significant effect on the expression of antiapoptotic markers in human melanoma cell lines.

Sec23a Inhibits The Self-Renewal of Melanoma Cancer Stem Cells via Inactivation of ER-Phagy

Background The genesis and developments of solid tumors, analogous to the renewal of healthy tissues, are driven by a subpopulation of dedicated stem cells, known as cancer stem cells (CSCs), that exhibit long-tern clonal repopulation and self-renewal capacity. CSCs may regulate tumor initiation, growth, dormancy, metastasis, recurrence and chemoresistance. While autophagy has been proposed as a regulator of the stemness of CSCs, the underlying mechanisms requires further elucidation. Methods The subpopulation of CSCs in human melanoma cell line M14 was established by repetitive enrichments for cells that consistently display anchorage-independent spheroid growth. The stemness properties of the CSCs were confirmed in vitro by the expressions of stemness marker genes, the single-cell cloning assay and the serial spheroid formation assay. Subcutaneous tumor transplantation assay in BALB/c nude mice was performed to test the stemness properties of the CSCs in vivo. The autophagic activity in cells was confirmed by the protein level of LC3 and P62, mRFP-LC3B punta and cytoplasmic accumulation of autolysosomes. The morphology of ER was detected with transmission electron microscopy. Results In the present study, by employing a stable CSC cell line derived from human melanoma cell line M14, we show for the first time that Sec23a inhibits the self-renewal of melanoma CSCs via inactivation of ER-phagy. Mechanistically, inhibition of Sec23a reduces ER stress and consequently FAM134B-induced ER-phagy. Furthermore, TCGA data mining and analysis show that Sec23a is a favorable diagnostic and prognostic marker for human skin cutaneous melanoma (SKCM). Conclusion Herein, this study has elucidated a new mechanism underlying the regulation of autophagy on stemness, i.e. CSCs can exploit the SEC23A/ER-stress/FAM134B/ER-phagy axis for the self-renewal. The results provide new ideas for comprehensive exploration of the regulatory network of CSC self-renewal and new potential targets for CSCs-based therapy strategies for malignant tumors.

An In Vitro-In Vivo Evaluation of the Antiproliferative and Antiangiogenic Effect of Flavone Apigenin against SK-MEL-24 Human Melanoma Cell Line

One of the most important class of natural compounds with successful preclinical results in the management of cancer is the flavonoids. Due to the plethora of biological activities, apigenin (4 ′ ,5,7 trihydroxyflavone) is a main representant of the flavone subclass. Although the antiproliferative and antiangiogenic effects of apigenin were studied on a significant number of human and murine melanoma cell lines, in order to complete the data existing in the literature, the aim of this study is to evaluate the in vitro effect of apigenin on SK-MEL-24 human melanoma cell line as well as in vivo on tumor angiogenesis using the aforementioned cell line on the chorioallantoic membrane assay. Results have shown that in the range of tested doses, the phytocompound presents significant antiproliferative, cytotoxic, and antimigratory potential at 30 μM, respectively, 60 μM. Moreover, the phytocompound in both tested concentrations limited melanoma cell growth and migration and induced a reduced angiogenic reaction limiting melanoma cell development.

Low expression of the PPARγ-regulated gene thioredoxin-interacting protein accompanies human melanoma progression and promotes experimental lung metastases

The thioredoxin system plays key roles in regulating cancer cell malignancy. Here we identify the Thioredoxin-interacting protein (TXNIP) as a gene, which expression is regulated by PPARγ in melanoma cells. We show that high TXNIP expression levels associate with benign melanocytic lesions, with tumor regression in patients on MAP kinase targeted therapy, with decreased proliferation in patients’ melanoma biopsies, and with cell cycle arrest in human melanoma cell lines. In contrast, reduced TXNIP expression associates with advanced melanoma and with disease progression in patients. TXNIP depletion in human melanoma cells altered the expression of integrin beta-3 and the localization of the integrin alpha-v/beta-3 dimer at their surface. Moreover, TXNIP depletion affected human melanoma cell motility and improved their capacity to colonize mouse lungs in an in vivo assay. This study establishes TXNIP as a PPARγ-regulated gene in melanoma cells, thereby suggesting a link between these two proteins both involved in the regulation of cancer and of energy metabolism. It also reveals that the decrease in TXNIP expression, which is observed in advanced patient tumors, likely favors lung metastatic seeding of malignant cells.

Quantitative Autofluorescence Imaging of A375 Human Melanoma Cell Samples: A Pilot Study

Introduction: Skin cancer is one of the most common types of malignancy worldwide. Human skin naturally contains several endogenous fluorophores, as potential sources can emit inherent fluorescence, called intrinsic autofluorescence (AF). The melanin endogenous fluorophore in the basal cell layer of the epidermis seems to have a strong autofluorescence signal among other ones in the skin. This pilot study aimed to investigate the feasibility of the detection of autofluorescence signals in the A375 human melanoma cell line in the cell culture stage using the FluoVision optical imaging system. Methods: The human skin melanoma cell line (A375) donated as a gift from Switzerland (University Hospital Basel) was cultured. For the imaging of the A375 human melanoma cell sample in this pilot study, the FluoVision optical imaging device (Tajhiz Afarinan Noori Parseh Co) was applied. The proposed clustering image processing code was developed based on the K-mean segmentation method, using MATLAB software (version 16). Results: The quantification of color pixels in the color bar along with the intensity score of the autofluorescence signal ranged between 0 and 70 was written in the image processing code execution and a threshold higher than 40%, proportional to the ratio of autofluorescent cells. The percentage of the signal of A375 autofluorescent melanoma cells in the 3 studied cell samples was calculated as 3.11%±0.6. Conclusion: This imaging method has the advantage of no need for fluorophore labels over the existing fluorescence imaging methods, and it can be regarded as one of the important choices of label-free imaging for this A375 melanoma cell line containing the intrinsic endogenous fluorophore in cell studies.