过表达Beclin1基因对横纹肌肉瘤RD细胞生物学行为的影响

Effects of Beclin1 Gene Overexpression on Biological Behavior of Rhabdomyosarcoma RD Cells

摘要

目的 本研究旨在探究Beclin1基因调控横纹肌肉瘤RD细胞的机制。方法 通过转染技术使Beclin1基因在RD细胞中过表达,将其分为实验组和阴性对照组。运用RT-qPCR及Western Blot分别从mRNA水平和蛋白水平检测Beclin1基因在RD细胞中的表达情况。采用CCK-8实验、细胞划痕愈合实验、Transwell实验以及TUNEL实验分别检测转染前后细胞的增殖、迁移、侵袭和凋亡情况。结果 实验组Beclin1基因的mRNA表达量为（4.06±0.46）,显著高于阴性对照组（P=0.008,P<0.01）,蛋白表达量为（1.84±0.30）,显著高于阴性对照组（P=0.032,P<0.05）;根据吸光度值绘制出实验组和阴性对照组的生长曲线,实验组细胞生长水平显著低于阴性对照组（P<0.05）;实验组细胞迁移能力明显弱于阴性对照组（P<0.01）,侵袭能力显著弱于阴性对照组（P<0.01）,细胞凋亡率显著高于阴性对照组（P<0.01）。结论 自噬调节基因Beclin1过表达可显著促进RD细胞凋亡,抑制RD细胞的增殖、迁移和侵袭能力。

Abstract

Objective The aim of this study was to investigate the mechanism by which Beclin1 gene regulates rhabdomyosarcoma RD cells. Methods Beclin1 gene was overexpressed in RD cells using the transfection technique and was divided into experimental group and negative control group. At the mRNA and protein levels, RT-qPCR and western blotting were used to detect the expression of Beclin1 gene, respectively. Cell proliferation, migration, invasion and apoptosis were detected based on CCK-8 test, scratch healing test, Transwell test and TUNEL test. Results Beclin1 gene had a significantly greater mRNA expression in the experimental group (4.06±0.46) compared to the negative control group (P=0.008, P<0.01). The expression level of protein in the experimental group was (1.84±0.30), which was higher than that in the negative control group (P=0.032, P<0.05). Two group's growth curve were drawn based on its optical density value. The experimental group's level of cell growth was lower than that of the negative control group (P<0.05). Cell migration in the experimental group was noticeably worse than that in the negative control group (P<0.01), and the experimental group's capacity for invasion was observably weaker than that of the negative control group (P<0.01). But the rate of apoptosis was notably higher than that of the negative control group (P<0.01). Conclusion The overexpression of autophagy regulatory gene Beclin1 can significantly promote the apoptosis of RD cells and inhibit the proliferation, migration and invasion of RD cells.

关键词

横纹肌肉瘤 / Beclin1 / 基因过表达 / 自噬

Key words

Rhabdomyosarcoma / Beclin1 / Gene overexpression / Autophagy

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基金资助

云南省教育厅科学研究基金项目（2024J0833）