靶向XBP1S的RNA干扰质粒对细胞增殖的影响

目的 构建靶向人XBP1S的siRNA真核表达载体（pSUPER-XBP1S）并观察其对人HeLa细胞和HepG2细胞增殖能力的影响.方法 设计并合成针对XBP1S基因的siRNA,退火成互补双链后克隆至真核表达载体pSUPER构建重组质粒,并将其转染入HeLa细胞和HepG2细胞中.采用RT-PCR检测转染前后XBP1S在HeLa细胞和HepG2细胞中的转录,Western印迹检测转染前后XBP1S蛋白的表达;MTT法、细胞计数检测重组质粒对HeLa细胞和HepG2细胞增殖能力的影响.结果 重组质粒能有效地抑制HeLa细胞和HepG2细胞中XBP1S基因的转录和表达;转染HeLa细胞和HepG2细胞后,细胞增殖抑制率及细胞增殖数与对照组比较,差异有统计学意义（P〈0.05）.结论 成功构建了靶向人XBP1S的siRNA表达载体pSUPER-XBP1S,并且有效的抑制了HeLa细胞和HepG2细胞中XBP1S的转录和表达,有效抑制了细胞的增殖能力.

Effects of siRNA Targeting-XBP1S on Cell Proliferation

Objective To construct siRNA eukaryotic expression vector targeting-XBP1S （pSUPER-XBP1S） and examine its effect on HeLa and HepG2 cells. Methods siRNA targeting- XBP1S segment was designed, synthesized and cloned into pSUPER vector to construct recombinant plasmid. The obtained plasmid was then transfected into human HeLa and HepG2 ceils. The expression of XBP1S gene in transfected HeLa and HepG2 cells was detected by RT-PCR and Western blotting assay. Cell proliferation was analyzed by MTT and cell counting. Results Recombinant pSUPER-XBP1S plasmid downregulated XBP1S at both mRNA and protein levels. Cell proliferation was significantly inhibited both with the difference being significant compared to control group （P 〈 0. 05） . Conclusion We have successfully constructed targeting-XBP1S siRNA expression vector pSUPER-XBPIS. The recombinant plasmid effectively suppresses the expression of XBP1S mRNA and protein in HeLa and HepG2 cells, and inhibits the cell proliferation, which may be useful for further study on XBP1S.