天津医药

天津医药 ›› 2019, Vol. 47 ›› Issue (7): 692-696.doi: 10.11958/20182237

• 细胞与分子生物学 • 上一篇    下一篇

OCCLUDIN对非小细胞肺癌SPC-A1细胞增殖和凋亡的影响及其机制研究

袁乐永，王梅芳   

  1. 1湖北医药学院基础医学院免疫教研室（邮编442000）；2湖北医药学院附属太和医院呼吸内科
  • 收稿日期:2019-01-03 修回日期:2019-04-17 出版日期:2019-07-15 发布日期:2019-08-01
  • 通讯作者: 王梅芳 E-mail:wangmeifang1979@163.com

The effect and mechanism of OCCLUDIN on proliferation and apoptosis of SPC-A1 cells

YUAN Le-yong,WANG Mei-fang   

  1. 1 Department of Immunology, School of Basic Medical Sciences, Hubei University of Medical, Hubei 442000, China; 2 Department of Respiratory, the Affiliated Taihe Hospital of Hubei University of Medical
  • Received:2019-01-03 Revised:2019-04-17 Published:2019-07-15 Online:2019-08-01

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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 Western blot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 120 h 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 Cyt C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。

关键词: 癌, 非小细胞肺, 肿瘤细胞, 培养的, 闭锁蛋白, 磷酸肌醇3-激酶类, 细胞增殖, 细胞凋亡

Abstract: 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.

Key words: carcinoma, non-small-cell lung, tumor cells, cultured, occludin, phosphatidylinositol 3-kinases, cell proliferation, apoptosis