Effects Of Silencing The Expression Of HnRNPA2/B1 Gene Using Lentivirus Mediated ShRNA On The Chemotherapy Sensitivity Of Cervical Cancer Cells And The Underlying Mechanism

Objective: To analysis hnRNPA2/B1 gene on Colony Formation, Cell Migration, Cell Invasion on cervical cancer cell and the Growth of Cervical cancer cell and sensitivity to drugs and to explore the mechanism. Methods: Designed 4pairs of interference sequence containing hnRNPA2/B1 shRNA(short hairpin RNA,shRNA) and 1 pairs of interference sequence containing negative control. TO constructe the recombinant Lentivirus plasmid containing short hairpin RNA targeting hnRNPA2/B1 gene, then transfected into lines CaSki and Hela. qRT-PCR detects expression of mRNA level of hnRNPA2/B1, the expression level of hnRNPA2/B1 protein was detected by Western blot. cell biological function were detected respectively by Clone Formation assay, Scratch assay and Invasion assay.IC50 of Lobaplatin and Irinotecan Hydrochloride and effects of growth ability was determined by MTT assay. Apoptotic rate and cell cycle in Caski cells and Hela cells were measured by flow cytometry(FCM). qRT-PCR and Western blot detects related genes of Bax, Bcl-2, CDH1. Results: The Lentivirus vector containing shRNA hnRNPA2/B1 was successfully constructed and transfected into Caski cells and HeLa cells. qRT-PCR was used to detect fact that the third and the fourth shRNA significantly reduced the expression of hnRNPA2/B1 mRNA in Caski cells,interference efficiency of the fourth was(74.79±3.38)%, The second, the third and the four interference sequence significantly decreased expression of hnRNPA2/B1 mRNA in Hela cells and the interference efficiency of the fourth was(76.58±3.55)% by qRT-PCR; The first, second, third and fourth interference sequences were significantly decreased the expression of hnRNPA2/B1 protein in Caski cells by Western blot, the interference efficiency of the fourth was(85.74±6.52)%, The first, second, third andfourth interference sequences were significantly decreased the expression of hnRNPA2/B1 protein in Hela cells by Western blot,the interference efficiency of the fourth was(73.25±2.78)%; Compared with the control group, gene silencing group of Caski group, Hela group in the ability of Colony Formation, Cell Migration, Cell Invasion were significantly decreased(P<0.01); blank control group and negative control group, the difference was not statistically significant(P>0.05);Compared with the control group, the proliferation ability of gene silencing group of Caski cell, Hela cell was significantly inhibited(P<0.01), the difference of blank control group and negative control group had no significant difference(P>0.05); Compared with the control group, gene silencing group of Hela cell, Caski cell IC50 of lobaplatin and IC50 of irinotecan were significantly decreased(P<0.01), the difference of blank control group and negative control group had no significant difference(P>0.05);compared with the control group, the silencing group of Caski group, Hela cell of the number of G1 cells was significantly higher than the control group(P<0.01), the difference of blank control group and negative control group had no significant difference(P>0.05); compared with the control group, the silencing group of CaSki cell, HeLa cell of the apoptosis rate of lobaplatin, irinotecan respectively was significantly higher than he negative control group(P<0.01), the difference of blank control group and negative control group was not statistically significant(P>0.05);Compared with the control group, the silencing group of CaSki cell, HeLa cell of Bax, bcl-2 and CDH1 expression were not statistically significant(P>0.05).Conclusion: Silencing hnRNPA2B1 gene can reduce the cervical carcinoma cell clone formation and migration and invasion, inhibit the proliferation of cervical cancer cells,block cells in the G1 phase and enhance anticancer drug sensitivity of lobaplatin and irinotecan.