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 proliferation, migration, and invasion of liver cancer cells

摘要

摘要:
背景抑制异常活化的信号通路，可有效抑制肿瘤细胞的生长和扩散，这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 qRT-PCR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；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 blot检测细胞增殖、侵袭和迁移中相关蛋白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表达。上调PTPRM表达可逆转 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，抑制肝癌细胞增殖、迁移、侵袭以及细胞增殖特异性周期蛋白(Cyclin)-D1的表达(P＜0.05)。
结论沉默PTPRM可能促进miR-139-5p的表达上调，导致肝癌细胞的增殖、迁移和侵袭能力降低。

Abstract:
Background Inhibiting aberrantly activated signaling pathways can effectively inhibit the growth and proliferation of tumor cells, which provides a new idea for the treatment of liver cancer.
Objective To investigate the impact of circRNA-PTPRM on liver cancer cell proliferation, migration, and invasion.
Methods qRT-PCR was used to detect the expression levels of PTPRM and miR-139-5p in normal liver cell line HL-7702 and liver cancer cell lines HepG2, Hep3B, Huh-7. Bioinformatics and dual luciferase assays were used to verify the interaction between PTPRM and miR-139-5p. Cell proliferation was assessed using MTT assay, while Transwell experiments were used to evaluate cell migration and invasion. Western blotting was applied to analyze the protein expression related to cell proliferation and invasion.
Results PTPRM levels were higher and miR-139-5p levels were 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 could increase miR-139-5p levels and reduce hepatocellular carcinoma cell proliferation, migration, invasion, and Cyclin D1 expression.
Conclusion Silencing PTPRM potentially elevates miR-139-5p expression, resulting in decrease of proliferation, migration, and invasion of liver cancer cells.

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