LncRNA H19通过海绵miR-675和上调CDH13的表达促进肺癌的增殖、侵袭和上皮-间质转化
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国家自然科学基金项目(81703033);山东省医药卫生科技发展计划项目(2018WS008)


LncRNA H19 promoted the proliferation, invasion and EMT of lung cancer through sponge miR-675 and up-regulation of CDH13 expression
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    目的 探讨LncRNA H19对肺癌SPC-A1细胞增殖、侵袭和上皮-间质转化(EMT)的影响及其作用机制。方法 实时荧光定量PCR检测LncRNA H19、miR-675和CDH13在SPC-A1细胞、BEAS-2B细胞中的差异表达;siRNA H19转染SPC-A1细胞,通过MTT、Transwell及Western blot检测SPC-A1细胞的增殖、侵袭和EMT能力。miRanda软件和双荧光素酶报告基因实验分析LncRNA H19和miR-675之间的作用靶点和相关性。miR-675 inhibitor转染SPC-A1细胞,检测SPC-A1细胞的增殖、侵袭和EMT能力。检测上调LncRNA H19通过miR-675对SPC-A1细胞增殖、侵袭和EMT的影响。TargetScan软件和双荧光素酶报告基因实验分析miR-675和CDH13之间的作用靶点和相关性。siRNA CDH13转染SPC-A1细胞,检测SPC-A1细胞的增殖、侵袭和EMT能力。RT-qPCR和Western blot检测LncRNA H19通过miR-675对CDH13表达的影响。结果 与BEAS-2B细胞相比,SPC-A1细胞中LncRNA H19和CDH13表达上调,miR-675表达下调(P<0.05)。siRNA H19转染SPC-A1细胞明显抑制了细胞的增殖、侵袭与EMT;LncRNA H19靶向且负调控miR-675;上调LncRNA H19通过miR-675表达促进SPC-A1细胞恶性发展。miR-675靶向且负调控CDH13。siRNA CDH13转染SPC-A1细胞明显抑制了细胞的增殖、侵袭与EMT。LncRNA H19通过miR-675上调CDH13表达。结论 LncRNA H19通过海绵miR-675和上调CDH13表达促进了SPC-A1细胞增殖、侵袭和EMT。

    Abstract:

    Objective To investigate the effects of lncRNA H19 on proliferation, invasion and EMT of lung cancer SPC-A1 cells and its mechanism. Methods The differential expressions of LncRNA H19, miR-675 and CDH13 in SPC-A1 cells and BEAS-2B cells were detected by real-time quantitative PCR. SPC-A1 cells were transfected with siRNA H19, and the proliferation, invasion and EMT ability of SPC-A1 cells were detected by MTT, Transwell and Western blot assay. The target and correlation between LncRNA H19 and miR-675 were analyzed by miRanda and dual luciferase reporter gene assay. SPC-A1 cells were transfected with miR-675 inhibitor to detect the proliferation, invasion and EMT ability of SPC-A1 cells. The effects of upregulated LncRNA H19 on proliferation, invasion and EMT of SPC-A1 cells via miR-675 were detected. TargetScan and dual luciferase reporter assay were used to analyze the target and correlation between miR-675 and CDH13. SPC-A1 cells were transfected with siRNA CDH13 to detect the proliferation, invasion and EMT ability of SPC-A1 cells. The effect of LncRNA H19 on CDH13 expression through miR-675 was detected by RT-qPCR and Western blot. Results Compared to BEAS-2B cells, the expressions of LncRNA H19 and CDH13 were upregulated (P<0.01), the expression of miR-675 was down-regulated (P<0.01) in SPC-A1 cells. The proliferation, invasion and EMT of SPC-A1 cells were significantly inhibited by siRNA H19 transfection. LncRNA H19 targeted and negatively regulated miR-675. Upregulation of LncRNA H19 through miR-675 or down-regulation of miR-675 expression promoted the malignant development of SPC-A1 cells. miR-675 targeted and negatively regulated CDH13. The proliferation, invasion and EMT of SPC-A1 cells were significantly inhibited by siRNA CDH13 transfection. LncRNA H19 up-regulated the expression of CDH13 by miR-675. Conclusion LncRNA H19 promoted proliferation, invasion and EMT of SPC-A1 cells through spongy miR675 and upregulation of CDH13 expression.

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  • 在线发布日期: 2022-08-19
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