SERPINB7对乳腺癌细胞增殖、凋亡、迁移和侵袭的影响

Effects of SERPINB7 on proliferation, apoptosis, migration and invasion of breast cancer cells

目的 探讨丝氨酸蛋白酶抑制剂B7(SERPINB7)对乳腺癌细胞增殖、凋亡、迁移和侵袭的作用及其可能机制。方法 将人乳腺癌细胞系MCF-7分为sh-SERPINB7组、空白对照组(BC组)和阴性对照组(NC组);其中,sh-SERPINB7组和NC组分别转染SERPINB7 RNA干扰慢病毒和阴性对照慢病毒。采用qRT-PCR法检测SERPINB7 mRNA表达,CCK-8法、流式细胞术、细胞划痕实验和Transwell实验分别检测细胞增殖、凋亡、迁移和侵袭,Western blot法检测SERPINB7、Bax、Bcl-2、Snail、Slug、转化生长因子β(TGF-β)、Smad3和磷酸化Smad3(p-Smad3)的蛋白表达。结果 MCF-7细胞中SERPINB7蛋白表达高于人正常乳腺上皮细胞系MCF-10A(P<0.05)。与BC组和NC组相比,sh-SERPINB7组MCF-7细胞中SERPINB7 mRNA表达以及Bcl-2、Snail、Slug、TGF-β、p-Smad3蛋白表达降低,细胞凋亡率升高,细胞划痕愈合率和侵袭细胞数减少,Bax蛋白表达增加(P<0.05)。结论 SERPINB7高表达于乳腺癌细胞,敲低SERPINB7可能通过抑制TGF-β/Smad3信号通路抑制乳腺癌细胞的迁移、侵袭,并促进其凋亡。

Objective To investigate the effects and underlying mechanism of serine protease inhibitor B7(SERPINB7) on the proliferation, apoptosis, migration and invasion of breast cancer cells.Methods Human breast cancer cell line MCF-7 was divided into three groups of sh-SERPINB7(transfected with SERPINB7 RNA interference lentivirus),BC(without treatment) and NC(transfected with negative control lentivirus).The mRNA expression of SERPINB7 was measured by qRT-PCR.The proliferation, apoptosis, migration and invasion of MCF-7 cells were determined by CCK-8,flow cytometry, cell scratch and Transwell assays, respectively.The protein expressions of SERPINB7,Bax, Bcl-2,Snail, Slug, TGF-β,Smad3 and p-Smad3 were detected by Western blot.ResultsThe protein expression of SERPINB7 in MCF-7 cells was higher than that in human mammary epithelial cell line MCF-10A(P<0.05).Compared with groups of BC and NC,the mRNA expression of SERPINB7 and protein expressions of Bcl-2,Snail, Slug, TGF-β and p-Smad3 were downregulated, rate of cell apoptosis was increased, healing rate of cell scratch and number of invasive cells were decreased, and protein expression of Bax was upregulated in group sh-SERPINB7(P<0.05).Conclusion SERPINB7 is highly expressed in breast cancer cells.Knockdown of SERPINB7 may inhibit the migration and invasion and promote apoptosis of breast cancer cells by inhibiting TGF-β/Smad3 signaling pathway.