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山东大学学报 (医学版) ›› 2026, Vol. 64 ›› Issue (7): 1-10.doi: 10.6040/j.issn.1671-7554.0.2025.0890

• 基础医学 •    下一篇

piR-020829通过胰岛素样生长因子结合蛋白-3调控肝癌增殖的机制

徐静怡1,宋彦威1,王丽婧1,主余华1,马铭泽1,刘欣欣2   

  1. 1.山东第一医科大学附属省立医院感染性疾病科, 山东 济南 250021;2.山东第一医科大学附属省立医院消化内镜中心, 山东 济南 250021
  • 出版日期:2026-07-10 发布日期:2026-07-21
  • 通讯作者: 刘欣欣. E-mail:1017838633@qq.com马铭泽. E-mail:mavictoryming@163.com
  • 基金资助:
    山东省自然科学基金面上项目(ZR2020MH240);国家自然科学基金青年项目(82000579)

Mechanism of piR-020829 in regulating hepatocellular carcinoma proliferation via insulin-like growth factor-binding protein 3

XU Jingyi1, SONG Yanwei1, WANG Lijing1, ZHU Yuhua1, MA Mingze1, LIU Xinxin2   

  1. 1. Department of Infectious Diseases, Shandong Provincial Hospital Affiliated to Shandong First Medical University, Jinan 250021, Shandong, China;
    2. Digestive Endoscopy Center, Shandong Provincial Hospital Affiliated to Shandong First Medical University, Jinan 250021, Shandong, China
  • Online:2026-07-10 Published:2026-07-21

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摘要： 目的 探讨piR-020829调控原发性肝癌恶性增殖的作用及相关分子机制。 方法 采用实时定量PCR法检测肝癌和癌旁肝组织piR-020829表达水平的差异;采用Kaplan-Meier生存曲线分析piR-020829表达水平与肝癌患者总生存期、无进展生存期的关系;采用短发夹RNA(short hairpin RNA, shRNA)干扰技术敲低SNU387细胞中piR-020829的表达(敲低组),并以转染阴性对照载体的SNU387细胞为敲低对照组;采用piR-020829模拟物处理Hep3B细胞实现过表达(piR-020829过表达组),并以转染模拟物阴性对照的Hep3B细胞为过表达对照组。采用CCK-8法检测piR-020829敲低表达/过表达后对肝癌细胞SNU387/Hep3B增殖活力的影响;采用肝癌细胞裸鼠皮下移植瘤实验,将32只裸鼠均分为拮抗剂组(尾静脉注射piR-020829 Antagomir)、拮抗剂对照组(注射Antagomir NC)、模拟物组(注射piR-020829 mimic)和模拟物对照组(注射mimic NC);至实验结束,观察不同处理组对裸鼠皮下成瘤瘤体质量的影响;采用克隆形成实验检测piR-020829敲低表达/过表达后对肝癌细胞SNU387/Hep3B体外增殖能力的影响。 结果 相较于癌旁组织,piR-020829在肝癌组织表达水平升高。生存分析显示,piR-020829相对高表达的肝癌患者总生存时间、无进展生存时间较低表达患者缩短,提示piR-020829高表达肝癌患者预后差。体外实验证实,piR-020829敲低表达后导致肝癌细胞增殖活力下降,克隆形成数目减少(P<0.05);piR-020829-mimic处理后导致肝癌细胞增殖活性升高,克隆形成数目增加。裸鼠皮下移植瘤实验证实,piR-020829 Antagomir处理后导致肝癌细胞裸鼠皮下成瘤质量低于拮抗剂对照组;piR-020829-mimic处理后导致肝癌细胞裸鼠皮下成瘤瘤质量高于模拟物对照组。拮抗剂组裸鼠移植瘤体免疫组织化学染色显示,Ki-67阳性率低于拮抗剂对照组(P<0.05)。piR-020829敲低表达导致凋亡相关蛋白B细胞淋巴瘤-2(B-cell lymphoma-2, Bcl-2)、B细胞淋巴瘤-xl(B cell lymphoma-xl,Bcl-xl)表达下调,半胱天冬酶-3(cysteine-aspartic protease 3, Caspase3)表达上调(P<0.05);piR-020829敲低表达导致细胞周期蛋白依赖性激酶4(cyclin-dependent kinase 4, CDK4)、细胞周期蛋白D1(G1/S-specific cyclin-D1, Cyclin D1)表达下调(P<0.05);piR-020829敲低表达导致其靶分子胰岛素样生长因子结合蛋白-3(insulin-like growth factor-binding protein 3, IGFBP3)表达上调(P<0.05)。 结论 piR-020829 通过靶向抑制IGFBP3表达而促进肝癌细胞恶性增殖,该分子有望成为抗肝癌靶向治疗新的分子靶标。

关键词: Piwi相互作用RNA, 胰岛素样生长因子结合蛋白-3, 肝癌, 增殖, 凋亡

Abstract: Objective To investigate the role of piRNA-020829(piR-020829)in regulating malignant proliferation of hepatocellular carcinoma(HCC)and its underlying molecular mechanism. Methods Quantitative real-time PCR was used to detect the expression levels of piR-020829 in HCC tissues and adjacent non-tumor liver tissues. Kaplan-Meier survival analysis was performed to evaluate the correlation between piR-020829 expression and overall survival(OS)and progression-free survival(PFS)in HCC patients. Short hairpin RNA interference technology was employed to knock down piR-020829 expression in SNU387 cells(knockdown group), with SNU387 cells transfected with negative control vector serving as negative control group. piR-020829 mimic was used to overexpress piR-020829 in Hep3B cells(overexpression group), with Hep3B cells transfected with mimic negative control serving as overexpression negative control group. CCK-8 assay was used to detect cell proliferation viability in SNU387/Hep3B cells after piR-020829 knockdown or overexpression. For in vivo experiments, 32 nude mice were randomly divided into 4 groups: antagomir group(tail vein injection of piR-020829 Antagomir), antagomir control group(injection of Antagomir NC), mimic group(injection of piR-020829 mimic), and mimic control group(injection of mimic NC); tumor weight was observed in each group at the end of the experiment. Colony formation assay was used to detect the in vitro proliferative capacity of SNU387/Hep3B cells after piR-020829 knockdown or overexpression. Results Compared with adjacent normal tissues, the expression level of piR-020829 was significantly elevated in hepatocellular carcinoma(HCC)tissues. Survival analysis showed that HCC patients with relatively high expression of piR-020829 had significantly shorter overall survival and progression free survival than those with low expression, indicating that high piR-020829 expression predicts a poor prognosis in HCC patients. In vitro experiments confirmed that knockdown of piR-020829 led to a significant decrease in the proliferation viability and colony formation ability of HCC cells(P<0.05), while treatment with piR-020829 mimic increased HCC cell proliferation activity and colony formation. Nude mouse subcutaneous xenograft experiments demonstrated that treatment with piR-020829 Antagomir resulted in a significantly lower tumor weight compared with the antagomir control group, whereas treatment with piR-020829 mimic led to a significantly higher tumor weight compared with the mimic control group. Immunohistochemical staining of xenograft tumors from the antagomir group showed a significantly lower Ki-67 positive rate than that in the antagomir control group(P<0.05). Knockdown of piR-020829 downregulated the expression of the apoptosis-related proteins B-cell lymphoma-2(Bcl-2)and B-cell lymphoma-xl(Bcl-xl), and upregulated the expression of cysteine-aspartic protease 3(Caspase-3)(P<0.05). Knockdown of piR-020829 also downregulated the expression of the cell cycle-related proteins CDK4 and Cyclin D1(P<0.05). Furthermore, mechanistic investigation revealed that knockdown of piR-020829 significantly upregulated the expression of its target molecule insulin-like growth factor-binding protein 3(IGFBP3)(P<0.05). Conclusion piR-020829 promotes malignant proliferation of HCC cells by targeting and inhibiting IGFBP3 expression, and this molecule may serve as a novel molecular target for anti-HCC targeted therapy.

Key words: Piwi-interacting RNA, Insulin-like growth factor-binding protein 3, Hepatocellular carcinoma, Proliferation, Apoptosis

中图分类号: 

  • R735.7
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