SNRPA1 up-regulation of FSTL1 contributes to proliferation and apoptosis of hepatocellular carcinoma cells
Objective Investigating whether SNRPA1 can participate in and regulate HCC progression by regulating FSTL1.Methods The cell lines LO2,BEL-7404,BEL-7402,SMMC-7721,and Hep-G2 were cul-tured.RT-PCR and western blot were used to detect the expression levels of SNRPA1 mRNA and protein level in LO2,BEL-7404,BEL-7402,SMMC-7721 and Hep-G2 cell lines.SNRPA1 knockdown in BEL-7404 was per-formed using lentivirus vectors encoding short hairpin RNA (shRNA),and cells were then transfected with (or with-out) the FSTL1 overexpression vector plasmid.High-content screening (HCS),MTT,western blot and flow cytome-try were used to detect the cell proliferation and apoptosis.Results SNRPA1 expression of HCC lines was signifi-cantly increased compared to LO2 cells (P<0.05).The expression of SNRPA1 in HCC lines was significantly high-er than that in LO2 cells,and the expression of SNRPA1 in BEL-7404 cells was the highest among all HCC lines.SNRPA1 levels were significantly decreased after SNR-PA1 knockdown;FSTL1 levels were significantly increased in cells after FSTL1 overexpression (P<0.05).HCS and MTT results showed that compared with the SNRPA1 knockdown group,the proliferation of overexpressing FSTL1 group was significantly increased after 1d to 5 d,and the western blot results showed that the expression of Ki-67 in the FSTL1 overexpression group was significantly increased (P<0.05).The flow cytometry results showed that compared with the SNRPA1 knockdown group,the apoptosis in the FSTL1 overexpression group was significantly inhibited,and western blot result showed that the expression levels of Caspase3 and Bim in the FSTL1 overexpression group were significantly reduced,and the Bcl-2 level was significantly increased (P<0.05).Conclusion SNRPA1 can promote HCC proliferation and inhibit HCC apoptosis by up-regulating FSTL1 and then regulating the expression of Ki-67 and Caspase 3.
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