非小细胞肺癌细胞中上皮特异性黏附分子的表达及其对细胞增殖和侵袭能力的影响
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摘要
目的:探讨上皮特异性黏附分子(EpCAM)在非小细胞肺癌(NSCLC)细胞中的表达及其对细胞增殖和侵袭能力的影响。方法:采用qRT-PCR和Western blot法检测人正常支气管上皮BEAS-2B细胞和NSCLC H226、A549、PC-9细胞中EpCAM mRNA和蛋白的表达。构建EpCAM低表达的A549细胞系,CCK-8法检测EpCAM对细胞增殖能力的影响,细胞平板克隆实验和Transwell实验检测EpCAM对细胞克隆形成能力和侵袭能力的影响,Western blot法检测EpCAM对细胞中Ki-67和MMP-9蛋白表达的影响。结果:H226、A549、PC-9细胞中EpCAM mRNA和蛋白的表达水平均高于BEAS-2B细胞,表达水平依次为A549>PC-9>H226(P<0.05)。成功干扰A549细胞中EpCAM表达后,细胞的增殖、克隆形成和侵袭能力均减弱,且Ki-67和MMP-9蛋白的表达均降低,与对照组相比,差异均有统计学意义(P<0.05)。结论:EpCAM在NSCLC细胞中高表达;干扰其表达可能通过下调Ki-67和MMP-9表达抑制A549细胞的增殖和侵袭。
Aim:To investigate the expression of epithelial cell adhesion molecule(EpCAM) in non-small cell lung cancer(NSCLC) cells and its effects on cell proliferation and invasion.Methods:The expressions of EpCAM mRNA and protein in human bronchial epithelial BEAS-2 B cells and NSCLC H226,A549 and PC-9 cells were detected by qRT-PCR and Western blot.After construction of EpCAM low expression A549 cell line,the effect of EpCAM on cell proliferation was measured by CCK-8 assay,the effects of EpCAM on cell cloning and invasion abilities were tested by cell clone and Transwell experiments,and the effect of EpCAM on the expression of cell proliferation related protein Ki-67 and MMP-9 protein in cells was examed by Western blot.Results:The expression levels of EpCAM mRNA and protein in H226,A549 and PC-9 cells were significantly higher than that of BEAS-2 B cells,and the intensity was A549 > PC-9 > H226 in turn(P<0.05).After successfully interfering EpCAM expression in A549 cells,the abilities of cell proliferation,cloning and invasion were significantly reduced,and the expressions of Ki-67 and MMP-9 protein were also significantly decreased.Compared with the control group,the difference was statistically significant(P<0.05).Conclusion:EpCAM is highly expressed in NSCLC cells;interfering its expression may inhibit the proliferation and invasion of A549 cells by downregulating the expressions of Ki-67 and MMP-9.
Aim:To investigate the expression of epithelial cell adhesion molecule(EpCAM) in non-small cell lung cancer(NSCLC) cells and its effects on cell proliferation and invasion.Methods:The expressions of EpCAM mRNA and protein in human bronchial epithelial BEAS-2 B cells and NSCLC H226,A549 and PC-9 cells were detected by qRT-PCR and Western blot.After construction of EpCAM low expression A549 cell line,the effect of EpCAM on cell proliferation was measured by CCK-8 assay,the effects of EpCAM on cell cloning and invasion abilities were tested by cell clone and Transwell experiments,and the effect of EpCAM on the expression of cell proliferation related protein Ki-67 and MMP-9 protein in cells was examed by Western blot.Results:The expression levels of EpCAM mRNA and protein in H226,A549 and PC-9 cells were significantly higher than that of BEAS-2 B cells,and the intensity was A549 > PC-9 > H226 in turn(P<0.05).After successfully interfering EpCAM expression in A549 cells,the abilities of cell proliferation,cloning and invasion were significantly reduced,and the expressions of Ki-67 and MMP-9 protein were also significantly decreased.Compared with the control group,the difference was statistically significant(P<0.05).Conclusion:EpCAM is highly expressed in NSCLC cells;interfering its expression may inhibit the proliferation and invasion of A549 cells by downregulating the expressions of Ki-67 and MMP-9.
引文
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[15]刘芳,诸兰艳.脂联素在非小细胞肺癌组织中的表达及其与基质金属蛋白酶-9和肿瘤血管生成的关系[J].中南大学学报(医学版),2015,40(6):579
[16]KROEPIL F,DULIAN A,VALLBHMER D,et al.High Ep CAM expression is linked to proliferation and lauren classification in gastric cancer[J].BMC Res Notes,2013,6:253
[17]GAO JJ,LIU XE,YANG F,et al.By inhibiting Ras/Raf/ERK and MMP-9,knockdown of Ep CAM inhibits breast cancer cell growth and metastasis[J].Oncotarget,2015,6(29):27187
[2]SORIA JC,OHE Y,VANSTEENKISTE J,et al.Osimertinib in untreated EGFR-mutated advanced non-small-cell lung cancer[J].N Engl J Med,2018,378(2):113
[3]KIM JH,BAE JM,SONG YS,et al.Clinicopathologic,molecular,and prognostic implications of the loss of EPCAMexpression in colorectal carcinoma[J].Oncotarget,2016,7(12):13372
[4]CAMPOS AK,HOVING HD,ROSATI S,et al.Ep CAM expression in lymph node and bone metastases of prostate carcinoma:a pilot study[J].Int J Mol Sci,2016,17(10):1650
[5]DAI M,YUAN F,FU CQ,et al.Relationship between epithelial cell adhesion molecule(Ep CAM)overexpression and gastric cancer patients:a systematic review and metaanalysis[J].PLo S One,2017,12(4):e0175357
[6]毛建英,冯建中,盛陈卓娅,等.上皮细胞黏附分子在非小细胞肺癌组织中表达及其临床意义[J].中华实验外科杂志,2016,33(11):2576
[7]LIU ZD,ZHEN DZ,LI YS,et al.Clinical significance of Ep CAM in circulating tumor cells in the peripheral blood during adjuvant therapy for postoperative non-small cell lung cancer patients[J].Chest,2016,149(4):325
[8]ZHOU N,WANG H,LIU H,et al.MTA1-upregulated EpCAM is associated with metastatic behaviors and poor prognosis in lung cancer[J].J Exp Clin Cancer Res,2015,34:157
[9]ZHENG J,ZHAO L,WANG Y,et al.Clinicopathology of Ep CAM and EGFR in human epithelial ovarian carcinoma[J].Open Med(Wars),2017,12(1):39
[10]OHASHI R,KAWAHARA K,FUJII T,et al.Higher expression of Ep CAM is associated with poor clinical and pathological responses in breast cancer patients undergoing neoadjuvant chemotherapy[J].Pathol Int,2016,66(4):210
[11]张泽栋,周逢强,徐宏,等.siRNA抑制EPCAM基因表达对结肠癌HCT116细胞株增殖和侵袭的影响[J].中国现代普通外科进展,2015,18(5):348
[12]涂雪松,胡利霞,瞿广桥,等.靶向下调上皮细胞黏附分子对结直肠癌干细胞功能及药物敏感性的影响[J].中国临床药理学杂志,2018,34(1):34
[13]TAYAMA S,MOTOHARA T,NARANTUYA D,et al.The impact of Ep CAM expression on response to chemotherapy and clinical outcomes in patients with epithelial ovarian cancer[J].Oncotarget,2017,8(27):44312
[14]刘明东,余宗阳,赵忠全,等.PD-L1、Ki67预测非小细胞肺癌术后进展的研究[J].现代肿瘤医学,2017,25(6):885
[15]刘芳,诸兰艳.脂联素在非小细胞肺癌组织中的表达及其与基质金属蛋白酶-9和肿瘤血管生成的关系[J].中南大学学报(医学版),2015,40(6):579
[16]KROEPIL F,DULIAN A,VALLBHMER D,et al.High Ep CAM expression is linked to proliferation and lauren classification in gastric cancer[J].BMC Res Notes,2013,6:253
[17]GAO JJ,LIU XE,YANG F,et al.By inhibiting Ras/Raf/ERK and MMP-9,knockdown of Ep CAM inhibits breast cancer cell growth and metastasis[J].Oncotarget,2015,6(29):27187