circRNA-PTPRM靶向miR-139-5p调控肝癌细胞增殖、迁移和侵袭的研究

陈秀华; 吴健晖; 陈良; 陈小芳; 余婷; 许芝彬

doi:10.12435/j.issn.2095-5227.2024.119

circRNA-PTPRM靶向miR-139-5p调控肝癌细胞增殖、迁移和侵袭的研究

circRNA-PTPRM targeting miR-139-5p regulates the proliferation, migration, and invasion of liver cancer cells

摘要

摘要:
背景通过抑制异常活化的信号通路，可以有效抑制肿瘤细胞的生长和扩散，为肝癌治疗提供了新的思路。
目的探讨circRNA-PTPRM对肝癌细胞增殖、迁移和侵袭的作用及其机制。
方法使用qRT-PCR法检测正常肝细胞系HL-7702和肝癌细胞系HepG2、Hep3B、Huh-7中PTPRM和miR-139-5p的表达水平；通过生物信息学和双荧光素酶检测实验验证PTPRM和miR-139-5p的靶向关系；采用MTT 法检测肝癌细胞增殖水平；Transwell迁移及侵袭实验检测细胞横向迁移及侵袭能力，Western印迹检测细胞增殖及侵袭迁移中相关蛋白Cyclin-D1的表达。
结果与正常肝细胞HL-7702比较，肝癌细胞系HepG2、Hep3B和Huh-7中PTPRM表达水平升高(P＜0.05)，miR-139-5p表达水平降低(P＜0.05)。选取HepG2细胞系进行后续实验，生物信息学显示circRNA-PTPRM与miR-139-5p存在结合位点，双荧光素酶报告实验结果及PCR显示PTPRM靶向调控miR-139-5p的表达。上调PTPRM表达则逆转 miR-139-5p过表达对肝癌细胞增殖、迁移和侵袭的抑制作用(P＜0.05)；过表达PTPRM可抑制miR-139-5p表达，沉默PTPRM可促进miR-139-5p表达，且可抑制肝癌细胞增殖、迁移、侵袭和细胞增殖特异性周期蛋白(Cyclin) -D1的表达(P＜0.05)。
结论通过沉默PTPRM可能促进miR-139-5p的表达上调，可能导致肝癌细胞增殖、迁移和侵袭能力降低。

Abstract:
Background Hepatocellular carcinoma, a common malignancy, involves complex signaling pathways critical for understanding its molecular mechanisms. Inhibiting these pathways can impede tumor growth and spread, suggesting new treatment approaches.
Objective To investigate the impact of circRNA-PTPRM on liver cancer cell proliferation, migration, and invasion.
Method The study involved detecting PTPRM and miR-139-5p levels in liver cancer cells using qRT-PCR. Bioinformatics and dual luciferase assays verified the interaction between PTPRM and miR-139-5p. Cell proliferation was assessed using MTT assay, while Transwell experiments evaluated cell migration and invasion. Western blotting analyzed protein expression related to cell proliferation and invasion.
Result PTPRM levels were higher and miR-139-5p levels lower in HepG2, Hep3B, and Huh-7 cell lines compared to normal liver HL-7702 cells (P<0.05). HepG2 was used for further experiments. Bioinformatics and experimental data confirmed PTPRM's regulatory effect on miR-139-5p expression. PTPRM upregulation negated miR-139-5p's suppressive effects on liver cancer cell proliferation, migration, and invasion. Conversely, silencing PTPRM increased miR-139-5p levels and reduced hepatocellular carcinoma cell proliferation, migration, invasion, and Cyclin D1 expression.
Conclusion Silencing PTPRM potentially elevates miR-139-5p expression, correlating with reduced proliferation, migration, and invasion of liver cancer cells.

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