摘要
目的分析长链非编码RNA CCAT1(LncRNA CCAT1)在非小细胞肺癌细胞系中的表达及作用。方法实时荧光定量PCR(qRT-PCR)测定非小细胞肺癌细胞系HCC827、H1650、H1299、A549及正常肺上皮细胞系BEAS-2B中LncRNA CCAT1的表达;将A549细胞系分为CCAT1-siRNA组、CTR-siRNA组及Mock组,经LipofectamineTM3000分别转染pcDNA3.1-LncRNA-CCAT1-shRNA、pcDNA3.1-LncRNA-CCAT1-NC及空白质粒pcDNA3.1;MTT实验测定细胞增殖能力,细胞划痕实验和Transwell实验测定细胞迁移、侵袭能力,qRT-PCR和蛋白免疫印迹实验分别测定细胞中miR-152和成纤维细胞生长因子2(FGF2)蛋白表达水平。结果 LncRNA CCAT1在非小细胞肺癌细胞系HCC827、H1650、H1299、A549中的表达量高于BEAS-2B(均P<0.01)。MTT结果示,转染72及96 h后,CCAT1-siRNA组细胞增殖活力显著低于CTR-siRNA组和Mock组(均P<0.05)。CCAT1-siRNA组划痕愈合率低于CTR-siRNA组[(33.45±2.62)%vs.(52.73±4.17)%,P<0.05]。CCAT1-siRNA组侵袭细胞数少于CTR-siRNA组[(93.7±6.2)vs.(172.8±11.5),P<0.01)]。CCAT-siRNA组miR-152表达量高于CTR-siRNA组[(3.73±0.18)vs.(1.02±0.05),P<0.01)]。CCAT-siRNA组FGF2蛋白表达量低于CTR-siRNA组[(0.59±0.03)vs.(1.01±0.06),P<0.01)]。结论LncRNA CCAT1在非小细胞肺癌细胞系中高表达,LncRNA CCAT1沉默可抑制非小细胞肺癌细胞增殖和侵袭,其机制可能与miR-152表达上调和FGF2表达下调有关。
Objective To investigate expression and effect of LncRNA CCAT1 in nonsmall-cell lung carcinoma(NSCLC)cells.Methods Real-time quantitative polymerase chain reaction(qRT-PCR)was used to detect the expression of LncRNA CCAT1 in NSCLC cell lines HCC827,H1650,H1299 and A549 and normal lung cell line,BEAS-2 B.The A549 cell line was divided into CCAT1-siRNA group,CTR-siRNA group and Mock group,which was respectively transfected with pcDNA3.1-LncRNA-CCAT1-shRNA,pcDNA3.1-LncRNA-CCAT1-NC and pcDNA3.1 by LipofectamineTM3000.MTT assay was used to test the proliferation ability.Cell wound scratch and transwell assay were used to test the invasion ability.The expression level of miR-152 and FGF2 protein was measured by qRT-PCR and Western blotting,respectively.Results LncRNA CCAT1 expression level was significantly higher in NSCLC cell lines,HCC827,H1650,H1299 and A549,than in normal lung cell line,BEAS-2 B(P<0.05).MTT showed that A490 nm of CCAT1-siRNA group was significantly lower than that in CTR-siRNA group and Mock group.The wound healing rate of CCAT1-siRNA group was significantly lower than that in CTR-siRNA group[(33.45±2.62)%vs.(52.73±4.17)%,P<0.05].The invasive cell number of CCAT1-siRNA group was significantly smaller than that in CTR-siRNA group[(93.7±6.2) vs.(172.8±11.5),P<0.01].The expression level of miR-152 in CCAT-siRNA group was significantly higher than that in CTR-siRNA group[(3.73±0.18) vs.(1.02±0.05),P<0.01].The expression level of FGF2 protein was significantly lower than that in CTR-siRNA group[(0.59±0.03) vs.(1.01±0.06),P<0.01].Conclusion The expression of LncRNA CCAT1 is significantly increased in the nonsmall-cell lung carcinoma cell lines compared to the normal lung cell line.The proliferation and migration could be inhibited by knockdown of LncRNA CCAT1,which may be caused by lower FGF2 level induced by higher miR-152.
作者
陈忠仁
欧宗兴
王蕾
沈彬
梁海梅
杨绪莉
黄实仁
Chen Zhongren;Ou Zongxing;Wang Lei(Department of Respiratory,Central South University Xiangya School of MedicineAffliated Haikou Hospital,Haikou 570208,China)
出处
《华中科技大学学报（医学版）》
CAS
CSCD
北大核心
2019年第5期513-517,共5页
Acta Medicinae Universitatis Scientiae et Technologiae Huazhong
基金
海南省自然科学基金资助项目（No.818MS136)
