miR-199a-3p负调控CBX7影响肺癌细胞NCI-H460的生物学行为研究

目的:探讨mi R-199a-3p负调控CBX7影响肺癌细胞NCI-H460的生物学行为。方法:qRT-PCR法检测并比较肺癌组织、癌旁正常组织、肺癌细胞、正常肺上皮细胞中的mi R-199a-3p m RNA相对表达量。比较远处转移肺癌组织、未转移肺癌组织中mi R-199a-3p m RNA相对表达量。qRT-PCR法、Western Blot法检测并比较肺癌组织、癌旁正常组织中的CBX7 m RNA及蛋白的表达水平。荧光素酶活性法检测mi R-199a-3p与靶基因CBX7的结合。比较mi R-199a-3p模拟物转染组与阴性对照组的肺癌细胞中的CBX7 m RNA相对表达量及CBX7蛋白表达水平。CCK8实验检测mi R-199a-3p对肺癌细胞增殖的促进作用。Tranwell实验检测mi R-199a-3p对肺癌细胞侵袭与迁移能力的影响。结果:肺癌组织中mi R-199a-3p明显高于癌旁正常组织,发生远处转移的肺癌组织中mi R-199a-3p m RNA的表达量明显高于未发生转移的肺癌组织,差异有统计学意义(P<0.001)。肺癌组织中CBX7m RNA、CBX7蛋白表达水平均明显低于癌旁正常组织,差异有统计学意义(P<0.001)。荧光素酶活性法证实mi R-199a-3p可与靶基因CBX7结合抑制CBX7的表达。肺癌细胞中mi R-199a-3p m RNA的相对表达量明显高于正常肺上皮细胞,CBX7 m RNA相对表达量明显低于正常肺上皮细胞(P<0.05)。对于肺癌细胞,mi R-199a-3p模拟物转染组的CBX7 m RNA相对表达量及CBX7蛋白表达水平均明显低于阴性对照组(P<0.001)。CCK8实验证实mi R-199a-3p能够促进肺癌细胞的增殖,Tranwell实验证实mi R-199a-3p对肺癌细胞侵袭与迁移具有积极的促进作用。结论:mi R-199a-3p在肺癌的发生发展过程中发挥重要作用,能够通过抑制CBX7基因的表达,促进肺癌细胞的增殖、侵袭和转移。

Investigate the Effect of miR-199a-3p on the Biological Behavior of NCI-H460 Lung Cancer Cells by Negatively Regulating CBX7

ABSTRACT Objective: To explore the effect of miR-199a-3p on the biological behavior of NCI-H460 lung cancer cells by negatively regulating CBX7. Methods: The relative expressions of miR-199a-3p mRNA in lung cancer tissues, adjacent normal tissues, lung cancer cells and normal lung epithelial cells were detected and compared by qRT-PCR. The relative expression levels of miR-199a-3p mRNA in distant metastatic lung cancer tissues and non-metastatic lung cancer tissues were compared. The expression levels of CBX7 mRNA and protein in lung cancer tissues and adjacent normal tissues were detected by qRT-PCR and Western Blot. The binding of miR-199a-3p to target gene CBX7 was detected by luciferase activity. CBX7 mRNA expression levels and CBX7 protein expression levels in lung cancer cells transfected with miR-199a-3p mimics and the negative control group were compared. CCK8 assay detected the promoting effect of miR-199a-3p on the proliferation of lung cancer cells. Tranwell assay examined the effect of miR-199a-3p on invasion and migration of lung cancer cells. Results: miR-199a-3p in lung cancer tissues was significantly higher than that in adjacent normal tissues, and the expression level of miR-199a-3p mRNA in lung cancer tissues with distant metastasis was significantly higher than that in lung cancer tissues without metastasis, the difference was statistically significant(P<0.001). The expression levels of CBX7 mRNA and CBX7 protein in lung cancer tissues were significantly lower than those in adjacent normal tissues(P<0.001). Luciferase activity confirmed that miR-199a-3p could bind to target gene CBX7 and inhibit the expression of CBX7.The relative expression level of miR-199a-3p mRNA in lung cancer cells was significantly higher than that in normal lung epithelial cells, and the relative expression level of CBX7 mRNA was significantly lower than that in normal lung epithelial cells (P<0.05). For lung cancer cells, the relative mRNA expression level and CBX7 protein expression level of miR-199a-3p mimics transfected group were significantly lower than those of the negative control group(P<0.001). CCK8 assay confirmed that miR-199a-3p could promote the proliferation of lung cancer cells, and Tranwell assay confirmed that miR-199a-3p had a positive promoting effect on the invasion and migration of lung cancer cells. Conclusion: miR-199a-3p plays an important role in the occurrence and development of lung cancer, and can promote the proliferation, invasion and metastasis of lung cancer cells by inhibiting the expression of CBX7 gene.