LncRNA TCF7通过调控miR-29和激活JAK/STAT2信号通路促进肺癌细胞的恶性生物学行为研究

LncRNA TCF7 promotes malignant biological behavior of lung cancer cells by regulating miR-29 and activating JAK/STAT2 signaling pathway

目的 探讨LncRNA TCF7通过调控miR-29和激活JAK/STAT2信号通路促进肺癌细胞的恶性生物学行为及其机制.方法 qPCR检测TCF7和miR-29在肺癌组织和不同肺癌细胞株中的表达情况;分析TCF7和肺癌患者的临床病理资料之间的关联;双荧光素酶报告基因检测TCF7与miR-29之间的相互作用;MTT增殖实验和Transwell侵袭实验检测抑制TCF7后肺癌细胞的增殖和侵袭能力的变化情况以及过表达miR-29的表达后对肺癌细胞增殖和侵袭能力的恢复情况;Western blotting检测抑制TCF7后JAK/STAT2信号通路蛋白的表达情况;裸鼠体外成瘤实验检测TCF7对体内成瘤的影响.结果 与其他肺癌细胞株相比,A549细胞中TCF7表达最高,miR-29的表达水平最高;TCF7的表达与肺癌的病理分期相关以及淋巴结转移情况有关,随着分期越高,TCF7在肺癌组织中表达越高,淋巴结转移的患者中TCF7的表达也相对较高;双荧光素酶实验证实TCF7能与miR-29的靶点特异性结合,可以调控miR-29的表达与活性;抑制TCF7的表达后可以促进肺癌细胞的增殖和侵袭能力;同时抑制miR-29的表达水平过后,肺癌细胞的增殖和侵袭能力得到一定程度的恢复;抑制TCF7的表达后,JAK/STAT2信号通路被相应地激活;与NC组相比,TCF7-siRNA组移植瘤的平均肿瘤体积和质量均相应降低.结论 TCF7可以调控miR-29的表达通过JAK/STAT2信号通路影响肺癌细胞的增殖和侵袭能力.

Objective To investigate the malignant biological behavior and mechanism of LncRNA TCF7 in lung cancer cells by regulating miR-29 and activating JAK/STAT2 signaling pathway.Methods qPCR was used to detect the expression of TCF7 and miR-29 in lung cancer tissues and different lung cancer cell lines.The relationship between TCF7 and clinicopathological data of lung cancer patients was analyzed.The dual luciferase reporter assay was used to detect the interaction between TCF7 and miR-29.MTT proliferation assay and Transwell invasion assay was used to detect the proliferation and invasion of lung cancer cells after inhibition of TCF7,respectively,and to analyze the relevant recovery after the overexpression of miR-29.The expression of JAK/STAT2 signaling pathway protein was detected by Western Blotting after TCF7 inhibition.The effect of TCF7 on tumor formation in vivo was detected by in vitro tumor formation assay in nude mice.Results Compared with other lung cancer cell lines,A549 cells had the highest expression of TCF7 and miR-29.The expression of TCF7 was associated with the pathological stage of lung cancer and lymph node metastasis,in which TCF7 was positively correlated with cancer stage and lymph node metastasis.The dual luciferase assay confirmed that TCF7 can specifically bind to the target of miR-29,and regulate the expression and activity of miR-29.The inhibition of the expression of TCF7 can promote the proliferation and invasion of lung cancer cells.After inhibiting the expression level of miR-29,the proliferation and invasion ability of lung cancer cells were partly restored.After inhibiting the expression of TCF7,JAK/STAT2 signaling pathway was activated accordingly.Compared with the non-small carcinoma group,the average tumor volume and mass of the transplanted tumor in the TCF7-siRNA group were reduced.Conclusions TCF7 can regulate the expression of miR-29 and affect the proliferation and invasion of lung cancer cells through JAK/STAT2 signaling pathway.