| Objective (s):Non-small cell lung cancer (NSCLC) has a high incidence rate worldwide. As the first leading cause of cancerous death NSCLC brings so many challenges to clinical diagnosis and treatment, and the underling driving mechanisms of it still need to be clarified. Yunnan province is the right place with high incidence of NSCLC. It is urgent to explore the specific drving mechanisms. The dysfunction of spliceosome may cause the alternatively splicing of downstream genes, and then participate in the tumorigenesis. SMN complex is the key compounds for the spliceosome assembly,and many components of the SMN complex are becoming the targets of treatment of cancers. However, the Gemin6 gene coding protein, the role of which in NSCLC is still a mystery, is an important component of SMN complex. At present, it is confirmed that the other components of SMN complex are involved in malignant biological behaviors and even the adjustable autophagy activity of cancer cells.Meanwhile, traditional Chinese medicines that can regulate the tumor cells’autophagy activity are emerging out, and have become the hotspot of medication research in the NSCLC. Therefore, this study aims to explore the connection between Gemin6 and malignant biological behaviors at the cell and tissue level. We would like to acquire scientific evidence that GEMIN6 is involved in proliferation, migration and other malignant biological behavior of NSCLC by detecting the activity of cell proliferation, migration, cell cycle process and autophagy proteins. And to further investigate whether berberine and other Chinese medicine monomers could regulate NSCLC malignant processes via GEMIN6. Finally, we hope this study could provide a new target for the diagnosis and treatment for NSCLC patients of Yunnan province.Methods:Westernblotting was used to test basal expression of GEMIN6 among NSCLC cell lines; siRNA was trasfected into the A549 and H1975 cell lines to knockdown GEMIN6 expression, which was validated by westernblotting; The cell proliferation experiment(MTS), transwell experiment and flow cytometry were used to detect GEMIN6’s influence on NSCLC cell growth; Westernblotting was used to validate the expression level of autophagy marker proteins (Beclinl and LC3 - Ⅰ/Ⅱ); MTS was carried out to test the influence of berberine on A549 cell proliferation;Westernblotting was used to test the effect on GEMIN6 and Beclinl expression after the cells were treated with five types of traditional Chinese medicine monomers;Immunohistochemical and HE staining to detect GEMIN6’s expression among cancerous tissues and benign tissues, and we use the SPSS software to analysis the clinical relevance between GEMIN6 and NSCLC.Results:1. The predicting survival curve according to online survival database(Kaplan-Meier plotter) showed that high expression of GEMIN6 was correlated with shorter survival of lung cancer patients2. GEMIN6 expression in NSCLC cancer tissues from Yunnan province is higher than the control group, but it has little clinical relevance3. GEMIN6 could increase the capability of proliferation and migration of NSCLC4. GEMIN6 promote G1 - S phase transition of NSCLC5. GEMIN6 inhibit the autophagy activity of NSCLC6. In A549 cells, berberine and curcumin could increase GEMIN6 and Beclinl expression; dihydroartemisinin could decrease GEMIN6 exprssion and increase Beclinl expression slightly at the same time; salidroside could increase GEMIN6 expression but had no effect on Beclinl.Conclusion(s):By inducing the G1 - S phase transition in H1975 and A549 cells, GEMIN6 could promote the proliferation and migration ability in both cell lines, and inhibit the activity of autophagy in A549 cells. Dihydroartemisinin could decrease GEMIN6 exprssion and increase the autophagy level, and GEMIN6 may be its’ downstrem target; Even though berberine, curcumin and salidroside could change the GEMIN6 expression level to some extend in A549 cells, but the correlation between their biological function and changed expression of GEMIN6 is undefined. |
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