Sorafenib In Combination With 5- Nitrogen Impurity- 2 ’- Deoxy Cytidine Regulating Of Liver Cancer Cell Lines DNMT3B Gene Expression Research

Objective:(1) to study the sorafenib combined with 5- nitrogen impurity- 2 ’- deoxy cytidine effect extremely to human liver cell lines and DNMT3 B gene expression(2)to investigate the possible molecular mechanism of sorafenib regulation of mi R- 143,mi R- 145 effects on the liver cancer cells and the possible molecular mechanisms.Methods:(1) to human liver cell line BEL- 7404, SMMC- 7721 as the research object, single drug and combination, using thiazole blue colorimetry, wound healing experiment, Transwell method and flow cytometry respectively test on human liver cell line proliferation, migration, invasion, and apoptosis.(2) the protein imprinting technology in the detection of liver cancer cell lines DNMT3 B, p- Akt- Ser473 and ERK protein expression;(3) by real-time fluorescent quantitative PCR technique to detect DNMT3 B gene m RNA expression in liver cancer cell lines.(4) in human liver cell line BEL- 7404 and immortalized hepatocyte L02 as the research object, using real-time fluorescent quantitative PCR technique to detect liver cancer cell line BEL-7404 and immortalized hepatocyte L02 mi R- 143, mi R- 145 expression situation,detection of sorafenib action BEL- 7404 cell mi R- 143, mi R- 145 expression after the change.(5) by liposome transient transfection mi RNA mimics to BEL- 7404 cell,detection of cell biology character change situation, study the corresponding mi R-143, mi R- 145 in the mechanism of action of liver cancer. And through the detection of DNMT3 B protein expression protein imprinting technology.Results:(1) the sorafenib in combination with 5- nitrogen impurity- 2 ’- deoxy cytidine after liver cancer cells can significantly inhibit cell proliferation and the dose-response relationship, and their combination have synergy; Able to suppress liver cancer cell migration and invasion ability; Can promote the apoptosis of cancer cells.(2) single drug and combination can reduce DNMT3 B and p- Akt- Ser473 protein expression, the expression of ERK protein had no effect.(3) single medicine and the combination can cut the cancer cells after DNMT3 B gene m RNA expression.(4)Mi R- 143 and mi R- 145 expression in HCC BEL- 7404 cells are immortalized liver L02 reduce; After sorafenib role in liver cancer cells express elevated mi R- 143 and mi R- 143.(5) transfection mi RNA mimics the liver cancer cells can inhibit cell proliferation and invasive ability, promoting apoptosis, sorafenib by raising the expression of mi R- 143 and mi R- 143 and cut DNMT3 B protein expression.Conclusion:1. Sorafenib combined 5- Aza- function CdR liver cancer cell have synergy, two drug combination can have synergistic antitumor effect effectively, and may through the inhibition of PI3K/Akt signaling pathway to reduce the expression of DNMT3 B.2. Mi R- 143 and mi R- 145 plays similar to the role of tumor suppressor genes in liver cancer3. Sorafenib by raising HCC BEL- 7404 cells the expression of mi R- 143 and mi R-143 and DNMT3 B lowered its target protein expression and thus play a role of anti-tumor.