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中国癌症防治杂志

中国癌症防治杂志 ›› 2025, Vol. 17 ›› Issue (4): 483-489.doi: 10.3969/j.issn.1674-5671.2025.04.13

• 论著 • 上一篇    下一篇

lncRNA ATP2C2⁃AS1通过调控PI3K/Akt/mTOR信号通路对骨肉瘤细胞增殖、凋亡的影响

  

  1. 华中科技大学同济医学院附属梨园医院康复医学科；湖北民族大学附属民大医院老年医学科；430065 武汉 3.湖北中医药大学中医学院；华中科技大学同济医学院附属梨园医院创面修复外科
  • 出版日期:2025-08-25 发布日期:2025-09-12
  • 通讯作者: 邓海波 E-mail：dhb0376@163.com
  • 基金资助:
     华中科技大学同济医学院附属梨园医院院内科学研究基金项目（2024LYYYLCYJ0008）

Effects of lncRNA ATP2C2⁃AS1 on the proliferation and apoptosis of osteosarcoma cells by regulating PI3K/Akt/mTOR signaling pathway

  • Online:2025-08-25 Published:2025-09-12

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摘要： 目的 探讨长链非编码RNA（long non⁃coding RNA,lncRNA）ATP酶分泌途径钙离子转运蛋白2（ATPase secretory pathway Ca2+ transporting 2，ATP2C2）反义RNA1(ATP2C2 antisense RNA 1,ATP2C2⁃AS1)对骨肉瘤细胞增殖、凋亡的影响及其机制。方法 采用qRT⁃PCR对人正常成骨细胞（hFOB 1.19）和人骨肉瘤细胞系（Saos⁃2、U2OS、143B和HOS）中ATP2C2⁃AS1的表达水平进行定量分析。骨肉瘤细胞Saos⁃2分别转染ATP2C2⁃AS1 siRNA（si⁃ATP2C2⁃AS1）或其阴性对照（si⁃NC），同时转染ATP2C2⁃AS1过表达质粒（oe⁃ATP2C2 ⁃AS1）或其阴性对照（Vector）。评估转染效率后，向转染oe⁃ATP2C2⁃AS1或Vector的Saos⁃2细胞加入磷脂酰肌醇3⁃激酶（phosphatidylinositol 3⁃kinase，PI3K）抑制剂NVP⁃BEZ235。上述处理细胞增殖活性通过CCK⁃8法测定，细胞凋亡水平采用流式细胞术检测。Western blot实验分析凋亡相关蛋白（Bax、Bcl⁃2和Cleaved⁃caspase⁃3）以及PI3K/蛋白激酶B（protein kinase B，Akt)/哺乳动物雷帕霉素靶蛋白（mammalian target of rapamycin，mTOR）信号通路相关蛋白的表达水平。结果 与hFOB 1.19细胞相比，人骨肉瘤细胞Saos⁃2、U2OS、143B和HOS细胞系中ATP2C2⁃AS1表达水平均显著升高（均P<0.001），其中Saos⁃2细胞的表达量最高。沉默ATP2C2⁃AS1可导致Saos⁃2细胞增殖活性降低，细胞凋亡增加，下调PI3K（p85α）、Bcl⁃2蛋白表达及p⁃Akt/Akt与p⁃mTOR/mTOR蛋白比值，上调Bax和Cleaved⁃ caspase⁃3蛋白表达（均P<0.05）。相反，过表达ATP2C2⁃AS1提高Saos⁃2细胞增殖活性，抑制凋亡，上调PI3K（p85α）、Bcl⁃2蛋白表达及p⁃Akt/Akt与p⁃mTOR/mTOR蛋白比值，下调Bax和Cleaved⁃caspase⁃3蛋白表达（均P<0.05）。NVP⁃BEZ235干预显著抑制ATP2C2⁃AS1过表达对Saos⁃2细胞的增殖促进作用，并诱导细胞凋亡。结论 ATP2C2⁃AS1可能通过激活PI3K/Akt/mTOR信号通路促进骨肉瘤细胞增殖，抑制其凋亡。

关键词: 骨肉瘤, ATP酶分泌途径钙离子转运蛋白2, 长链非编码RNA, 增殖, 凋亡

Abstract: Objective To investigate the effects of the long non⁃coding RNA (lncRNA) ATPase secretory pathway Ca2+ transporting 2 antisense RNA 1 (ATP2C2⁃ AS1) on the proliferation andapoptosis of osteosarcoma cells， as well as to elucidate its underlying mechanism.  Methods The expression levels of ATP2C2⁃AS1 in human normal osteoblasts cell (hFOB 1.19) and human osteosarcoma cell lines (Saos⁃2， U2OS， 143B， and HOS) were quantified using qRT⁃PCR. Saos⁃2 osteosarcoma cells were transfected with either an ATP2C2⁃AS1 siRNA (si⁃ATP2C2⁃AS1) and its negative control (si⁃NC)， as well as ATP2C2⁃AS1 overexpression plasmid (oe⁃ATP2C2⁃AS1) or its negative control (Vector). Following the assessment of transfection efficiency， the PI3K inhibitor (NVP⁃BEZ235) was used to Saos⁃2 cells transfected with either the Vector or oe⁃ATP2C2⁃AS1. Cell proliferation activity was measured using the cell counting kit⁃8 (CCK⁃8) method， while apoptosis levels were assessed via flow cytometry. Western blot was used to detect the expression levels of apoptosis⁃related proteins (Bax， Bcl⁃2， and Cleaved⁃caspase⁃3) and proteins associated with the phosphatidylinositol 3⁃kinase (PI3K)/protein kinase B (Akt)/mammalian target of rapamycin (mTOR) signaling pathway. Results Compared with hFOB 1.19 cells， the expression levels of ATP2C2⁃AS1 were markedly elevated in Saos⁃2， U2OS， 143B and HOS cell lines(all P<0.001)， with the most pronounced expression observed in Saos⁃2 cells. The silencing of ATP2C2⁃AS1 reduced the proliferation activity in Saos⁃2 cells， an increase cell apoptosis， and a downregulation of PI3K (p85α)， Bcl⁃2 protein， and the p⁃Akt/Akt and p⁃mTOR/ mTOR proteins ratios， while it upregulated the expression of Bax and Cleaved⁃caspase⁃3 proteins (all P<0.05). Conversely， the overexpression of ATP2C2⁃AS1 enhanced the proliferation activity of Saos⁃2 cells， inhibited  apoptosis， and led to an upregulation of PI3K (p85α)， Bcl⁃2 protein， and the ratios of p⁃Akt/Akt and p⁃mTOR/mTOR protein ratios， while downregulating the expression of Bax and Cleaved⁃ caspase⁃3 proteins (all P<0.05). The intervention with NVP⁃BEZ235 significantly counteracted the proliferation effects of  ATP2C2⁃AS1 overexpression in Saos⁃2 cells， and induced apoptosis. Conclusions ATP2C2⁃AS1 may promote the proliferation and inhibit the apoptosis of osteosarcoma cells through the activation of the PI3K/Akt/mTOR signaling pathway.

Key words: Osteosarcoma, ATPase secretory pathway Ca2+ transporting 2, Long non?coding RNA, Proliferation, Apoptosis

中图分类号: 

  • R738.1