脂联素受体AdipoR1对肺癌细胞PC9增殖和迁移的影响
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国家自然科学基金青年基金(81601189);山东省自然科学基金(ZR201702210055,ZR2014HQ080,ZR201709220399,ZR2019MC026);山东省医药卫生科技发展计划项目(2017WS154);滨州医学院科研计划与科研启动基金(BY2015KJ10)


The effects of adiponectin receptor AdipoR1 on proliferation and migration of PC9 cells
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    目的:观察过表达或敲低人脂联素受体1(AdipoR1)对肺癌细胞PC9增殖和迁移的影响,阐明AdipoR1对肺癌的调控作用。方法:逆转录聚合酶链反应(RT-PCR)检测AdipoR1在PC9细胞及正常人支气管上皮细胞HBEC中的表达量;利用分子克隆的方法构建AdipoR1过表达和shRNA干扰载体,转染至PC9细胞中并采用RT-PCR和Western blot方法检测AdipoR1的表达情况。运用CCK8方法和划痕愈合实验检测AdipoR1激动剂AdipoRon、AdipoR1过表达及AdipoR1敲低对PC9细胞的增殖和迁移活性的影响。结果:RT-PCR结果显示AdipoR1在PC9细胞中的表达量明显降低(P < 0.05)。RT-PCR和Western blot结果显示AdipoR1过表达的PC9细胞中AdipoR1 mRNA和蛋白的表达量明显高于空载体细胞(P均<0.05),AdipoR1干扰组AdipoR1表达量明显低于阴性对照细胞(P均<0.05)。CCK8和划痕愈合实验结果显示,在24 h和48 h时,与溶剂对照组相比,AdipoRon能显著降低PC9细胞的增殖和划痕愈合率,过表达AdipoR1的PC9细胞增殖和划痕愈合率明显低于空载体细胞,而AdipoR1敲低能明显促进PC9细胞的增殖和划痕愈合率(P均<0.05)。结论:AdipoR1激动剂AdipoRon或AdipoR1过表达能抑制肺癌细胞PC9的增殖和迁移活性,而AdipoR1敲低则能明显促进肺癌细胞PC9的增殖和迁移。

    Abstract:

    Objective:To observe the effects of human adiponectin receptor(AdipoR1)gene overexpression and knockdown on PC9 cells proliferation and migration,and to clarify the regulatory effect of AdipoR1 on lung cancer. Methods:The expression levels of AdipoR1 in PC9 cells and control HBEC were deteceted by RT-PCR. The overexpression vector and shRNA interference vector of AdipoR1 were established by molecular clone method and transfected into PC9 cells,the expression levels of AdipoR1 mRNA and protein were detected by RT-PCR and Western blot. CCK8 method and scratch healing experiment were used to detect the effect of AdipoR1 agonist AdipoRon,AdipoR1 overexpression,and AdipoR1 knockdown on proliferation and migration ability of PC9 cells. Results:RT-PCR results showed that the expression of AdipoR1 was decreased in PC9 cells(P < 0.05). RT-PCR and Western blot results showed that mRNA and protein expression levels of AdipoR1 were up-regulated in AdipoR1 overexpressed PC9 cells(P < 0.05),and down-regulated in AdipoR1 knockdown PC9 cells(P < 0.05). CCK8 and scratch healing results showed that the proliferation and healing percentage of PC9 cells treated by AdipoRon for 24 or 48 h were significantly decreased when compared with the vehicle group(all P< 0.05). The proliferation and healing percentage were also significantly lower in AdipoR1 overexpression PC9 cells and higher in AdipoR1 knockdown PC9 cells than theircontrols after cultured for 24 or 48 h(all P < 0.05). Conclusion:AdipoR1 agonist AdipoRon,AdipoR1 overexpression can inhibit the proliferation and migration ability of lung cancer PC9 cells,while AdipoR1 knockdown can increase the proliferation and migration ability of lung cancer PC9 cells.

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代娟娟,杨丽娟,武 艳,杜 静,孟凡涛,安佳佳,李 晨.脂联素受体AdipoR1对肺癌细胞PC9增殖和迁移的影响[J].南京医科大学学报(自然科学版),2020,(1):15-20

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  • 收稿日期:2019-05-30
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  • 在线发布日期: 2020-02-09
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