摘要
目的探讨微小核糖核酸(miRNA,miR)-187在非小细胞肺癌(NSCLC)中表达的变化对癌细胞功能和吉非替尼耐药性的影响。方法选择来源于江苏省人民医院30例原发性NSCLC手术切除患者的肺癌组织和相应的癌旁组织标本。利用qRT-PCR检测肺癌组织和相应癌旁正常组织标本中的miR-187水平;CCK8法检测肺腺癌PC9细胞株和肺腺癌吉非替尼耐药株PC9G的细胞增殖活性、凋亡能力;克隆形成实验测定细胞增殖能力;Transwell实验测定细胞侵袭能力;荧光素酶报告基因验证miR-187与下游基因[细胞间黏附分子(ICAM1)]的结合;Western blot法用于验证miR-187下游蛋白的表达情况。结果相比癌旁正常组织,miR-187在原发NSCLC组织中明显下调;且在吉非替尼耐药株PC9G中,miR-187表达量明显低于吉非替尼敏感的PC9细胞(P<0.01)。过表达miR-187可以抑制PC9G细胞的增殖和迁移,逆转其对吉非替尼的耐药性。相反,在PC9细胞中沉默miR-187能够显著增加PC9细胞的增殖和侵袭性,并赋予其对吉非替尼的耐药性。另外,实验证实,ICAM1基因含有的miR-187-5p同源位点(3′-UTR的UGUAGC),可能为miR-187的靶基因。结论miR-187能够抑制NSCLC细胞的增殖,促进肺癌细胞凋亡,且可以通过调节ICAM1逆转PC9G细胞对吉非替尼的耐药性。
Objective To investigate the effect of the expression of microRNA( miRNA,miR)-187 on the function of lung cancer cells and gefitinib resistance in non-small cell lung cancer ( NSCLC). Methods Lung cancer tissues and corresponding paracancerous tissues from 30 NSCLC patients with surgical resection in Jiangsu Province Hospital were collected. The levels of miR-187 in two kinds of specimens were detected by qRT-PCR;the proliferative activity and apoptosis of PC9 and PC9G ( PC9-gefitinib resistant cells) cells were detected by CCK8 method;colony formation assay and transwell invasion assay were used to determine the ability of cell proliferation and invasion;the luciferase reporter system was used to verify the binding of miR-187 to downstream genes [ intercellular adhesion molecule( ICAM1 )];Western blot was used to detect the expression of downstream protein of miR-187. Results Compared with normal tissues adjacent to the tumor,miR-187 significantly decreased in primary NSCLC tissues,and its expression in gefitinib-resistant PC9G cells was significantly lower than that in gefitinib-sensitive PC9 cells ( P < 0. 01 ). Overexpression of miR-187 can inhibit the proliferation and migration of PC9G cells and reverse their resistance to gefitinib. In contrast, silencing miR-187 in PC9 cells significantly increased the proliferation and invasiveness of PC9 cells and endowed them with gefitinib resistance. In addition,ICAM1 gene contains a homology of miR-187-5p,which may be the target gene of miR-187. Conclusion miR- 187 can inhibit the proliferation of NSCLC cells, promote the apoptosis of lung cancer cells and reverse the resistance of PC9G cells to gefitinib by regulating ICAM1.
作者
马浩
孙立柱
丁昕
渠德宝
章龙珍
MA Hao;SUN Li-zhu;DING Xin;QU De-bao;ZHANG Long-zhen(Xuzhou Medical University,Xuzhou,Jiangsu 221004,China)
出处
《中国临床研究》
CAS
2019年第10期1336-1341,共6页
Chinese Journal of Clinical Research
