扣针蛋白5(FBLN5)在肺癌组织中表达及对肺癌细胞增殖、迁移及侵袭的影响

The expression of fibulin 5(FBLN5)in lung cancer tissue and its effects on the proliferation and migration of lung cancer cells

目的:探讨扣针蛋白5(FBLN5)在肺癌组织中的表达及其对肺癌细胞(HCC827)增殖、迁移和侵袭的影响。方法:选取2019年1月至2020年6月在石家庄市人民医院行手术切除的67例肺癌组织与癌旁组织,采用实时荧光定量PCR(RT-qPCR)检测肺癌组织及癌旁组织FBLN5 mRNA表达水平,分析FBLN5表达与肺癌临床病理特征的相关性;体外培养肺癌细胞系HCC827细胞及BEAS-2B细胞,将HCC827细胞分为空白对照组(Control组)、FBLN5过表达组(pcDNA-FBLN5组)和阴性对照组(pcDNA-NC组)。RT-qPCR法检测FBLN5 mRNA表达;MTT法与Edu染色检测细胞增殖;Transwell小室实验检测细胞迁移与侵袭;免疫蛋白印迹法(Western blotting)检测细胞中Ki67、PCNA、MMP-2、MMP-9蛋白表达。结果:FBLN5 mRNA在肺癌组织中表达水平(0.41±0.07)低于癌旁组织的(1.00±0.00)(P<0.05);肺癌组织中FBLN5表达与TNM分期和淋巴结转移有关(P<0.05);HCC827细胞FBLN5 mRNA及蛋白表达水平低于BEAS-2B细胞(P<0.05);与Control组和pcDNA-NC组相比,pcDNA-FBLN5组HCC827细胞FBLN5 mRNA及蛋白表达水平升高(P<0.05),细胞增殖活性、迁移与侵袭细胞数目及细胞中Ki67、PCNA、MMP-2、MMP-9蛋白表达水平降低(P<0.05)。结论:FBLN5在肺癌组织中表达下调,且与肺癌的发生发展相关,过表达FBLN5可显著抑制肺癌细胞增殖、迁移及侵袭。

Objective:To investigate the expression of fibulin 5(FBLN5)in lung cancer and its effects on the proliferation,migration,and invasion of lung cancer cells(HCC827).Methods:From January 2019 to June 2020,67 surgical excision specimens of lung cancer tissue and adjacent tissue in our hospital were selected.realtime reverse transcription-quantitative PCR(RT-qPCR)was used to detect the expression level of FBLN5 mRNA in lung cancer tissue and adjacent tissue.The correlation between FBLN5 expression and clinicopathological characteristics of lung cancer patients was analyzed.Lung cancer cells(cell lines HCC827 and BEAS-2B)were cultured in vitro,and HCC827 cells were divided into blank control group(control group),FBLN5-overexpression group(pcDNA-FBLN5 group),and negative control group(pcDNA-NC group).The RT-qPCR method was used to detect FBLN5 mRNA expression.MTT and Edu staining methods were used to detect cell proliferation.A transwell chamber experiment was used to detect cell migration and invasion.Western blotting(WB)was used to detect the expression of Ki67,PCNA,MMP-2,and MMP-9 proteins in the cells.Results:The expression level of FBLN5 mRNA in lung cancer tissue was 0.41±0.07,which was lower than that in adjacent tissue(1.00±0.00)(P<0.05).The expression of FBLN5 in lung cancer tissue was related to TNM staging and lymph node metastasis(P<0.05).The expression levels of FBLN5 mRNA and protein in HCC827 cells were significantly lower than that in BEAS-2B cells(P<0.05).Compared with the control group or the pcDNA-NC group,the FBLN5 mRNA and protein expression levels of HCC827 cells in the pcDNA-FBLN5 group were significantly higher(P<0.05);the less visible cell proliferation activity,the decreased numbers of migration and invasion cells,and the significantly lower expression levels of Ki67,PCNA,MMP-2,and MMP-9 proteins of HCC827 cells were also detected in the pcDNA-FBLN5 group rather than the control group or the pcDNA-NC group(P<0.05).Conclusion:FBLN5 is down-regulated in lung cancer and is related to the occurrence and development of lung cancer.Overexpression of FBLN5 can significantly inhibit the proliferation,migration,and invasion of lung cancer cells.