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目的:探讨靶向B7-H4基因的小干扰RNA(small interference RNA,siRNA)对人肺腺癌细胞A549增殖、侵袭和迁移的影响。方法:体外化学合成B7-H4特异性siRNA(B7-H4-siRNA),转染A549细胞,MTT法、流式细胞术、Western blotting分别检测A549细胞的增殖、细胞周期,以及B7-H4和Cyclin D1蛋白的水平;Transwell实验检测A549细胞体外侵袭和迁移能力。结果:B7-H4-siRNA成功转染入A549细胞,Western blotting结果显示,B7-H4-siRNA转染抑制A549细胞中B7-H4和Cy-clin D1蛋白的表达。B7-H4-siRNA转染后A549细胞增殖活性明显下降,B7-H4-siRNA组A549细胞的倍增时间明显长于未转染组、Ctrl-siRNA组和转染试剂对照组[(33.78±0.26)h vs(28.69±0.18)、(27.32±0.13)、(26.93±0.19)h,P<0.05]。B7-H4-siRNA转染使A549细胞阻滞在G1期。B7-H4-siRNA组A549细胞的侵袭细胞数明显少于未转染组[(89.80±0.99)个vs(186.20±1.33)个,P<0.05]。B7-H4-siRNA组A549细胞的迁移细胞数明显少于未转染组、Ctrl-siRNA组和转染试剂对照组[(60.20±0.37)个vs(102.57±0.52)、(100.72±0.31)、(98.65±0.21)个,P<0.05〗。结论:B7-H4-siRNA能沉默A549细胞中B7-H4的表达,抑制A549细胞的增殖、侵袭和迁移,B7-H4可能成为肺癌基因治疗的候选靶点。
Objective: To investigate the effect of small interfering RNA (siRNA) targeting B7-H4 gene on the proliferation, invasion and migration of human lung adenocarcinoma A549 cells. Methods: B7-H4 specific siRNA (B7-H4-siRNA) was synthesized and transfected into A549 cells in vitro. The proliferation and cell cycle of A549 cells were detected by MTT assay, flow cytometry and Western blotting respectively. D1 protein level; Transwell assay A549 cell invasion and migration in vitro. Results: B7-H4-siRNA was successfully transfected into A549 cells. The results of Western blotting showed that B7-H4-siRNA transfection inhibited the expression of B7-H4 and Cy-clin D1 in A549 cells. The proliferation activity of A549 cells was significantly decreased after B7-H4-siRNA transfection. The doubling time of A549 cells in B7-H4-siRNA group was significantly longer than that of untransfected A549 cells [(33.78 ± 0.26) h vs (28.69 ± 0.18), (27.32 ± 0.13), (26.93 ± 0.19) h respectively, P <0.05]. A549 cells were arrested in G1 phase by B7-H4-siRNA transfection. The number of invasive cells in B7-H4-siRNA A549 cells was significantly less than that in untransfected cells [(89.80 ± 0.99) vs (186.20 ± 1.33), P <0.05]. The number of migrating cells in A549 cells in B7-H4-siRNA group was significantly lower than that in untransfected cells (60.20 ± 0.37 vs 102.57 ± 0.52, 100.72 ± 0.31, (98.65 ± 0.21), P <0.05. CONCLUSION: B7-H4-siRNA can silence the expression of B7-H4 in A549 cells and inhibit the proliferation, invasion and migration of A549 cells. B7-H4 may be a potential target for gene therapy of lung cancer.