摘要
目的探讨OCCLUDIN对非小细胞肺癌细胞SPC-A1增殖和凋亡的影响及其作用机制。方法培养非小细胞肺癌细胞SPC-A1,通过siRNA转染干扰OCCLUDIN表达,实验中分为空白对照组(control组)、空转组(siCtrl组)和siRNA转染组(siOCLN组)。采用CCK-8、流式细胞术和Westernblot检测OCCLUDIN对SPC-A1细胞增殖和细胞凋亡的影响。结果siOCLN组细胞在24、48、72、96和120h的增殖能力较control组和siCtrl组均明显降低(P<0.05)。siOCLN组细胞凋亡率高于control组和siCtrl组(P<0.05);siOCLN组Bax、caspase-3、caspase-9、AIF和CytC等凋亡相关蛋白水平较control组和siCtrl组表达明显升高(P<0.05),而抗凋亡蛋白Bcl-2水平较control组和siCtrl组表达明显下降(P<0.05)。同时,siOCLN组AKT和PI3K蛋白磷酸化水平较control组和siCtrl组明显降低(P<0.05)。结论OCCLUDIN可能通过调控PI3K/AKT信号通路促进SPC-A1细胞的增殖和抑制凋亡。
Objective To investigate the effect of OCCLUDIN on proliferation and apoptosis of SPC-A1 cells and its mechanism thereof. Methods Non-small cell lung cancer cell line SPC-A1 was cultured and interfered with OCCLUDIN expression by siRNA transfection. The cells were divided into blank control group (control group), idling group (siCtrl group) and siRNA transfection group (siOCLN group). The effects of OCCLUDIN on proliferation and apoptosis of SPC-A1 cells were detected by CCK-8 assay, flow cytometry and Western blot assay. Results The cell proliferation abilities of siOCLN cells were significantly lower at 24 h, 48 h, 72 h, 96 h and 120 h than those of control group and siCtrl group (P<0.05). The apoptosis rate was significantly higher in siOCLN group than that of control group and siCtrl group (P<0.05). The levels of apoptosis-related proteins such as Bax, caspase-3, caspase-9, AIF and Cyt C were significantly higher in siOCLN group than those in control group and siCtrl group (P<0.05). The level of anti-apoptotic protein Bcl-2 was significantly lower in siOCLN group than that of control group and siCtrl group (P<0.05). At the same time, the phosphorylation levels of AKT and PI3K protein were significantly lower in siOCLN group than those in control group and siCtrl group (P<0 .05). Conclusion OCCLUDIN may promote the proliferation and inhibit apoptosis of SPC-A1 cells by regulating PI3K/AKT signaling pathway.
作者
袁乐永
王梅芳
YUAN Le-yong;WANG Mei-fang(Department of Immunology,School of Basic Medical Sciences,Hubei University of Medical,Hubei 442000,China;Department of Respiratory,the Affiliated Taihe Hospital of Hubei University of Medical)
出处
《天津医药》
CAS
北大核心
2019年第7期692-696,共5页
Tianjin Medical Journal
基金
国家自然科学基金资助项目(81802090)
湖北省自然科学基金资助项目(2017CFB123)
