摘要
为探讨长链非编码RNA(long non-coding RNA, lncRNA)浆细胞瘤变体易位1(plasmacytoma variant translocation 1, PVT1)对宫颈癌细胞增殖、凋亡、迁移、侵袭的影响,和其对miR-484、 Krüppel样因子12(Krüppel-like factor 12, KLF12)的调控,以及其对IL-6/STAT3信号的调节机制,采用qRT-PCR检测宫颈癌组织和宫颈癌细胞系中lncRNA PVT1、 miR-484以及KLF12 mRNA的表达;双荧光素酶试验检测lncRNA PVT1、 miR-484、 KLF12的关系;以Sh-PVT1、 Sh-PVT1+antagomir、 Sh-PVT1+antagomir+pcDNA3.1转染细胞,检测细胞的增殖、凋亡、迁移以及侵袭性能;检测肿瘤细胞的体内生长与转移性能;Western blotting检测各组细胞和小鼠肿瘤组织中KLF12、 B淋巴细胞瘤2(B-cell lymphoma 2, Bcl-2)和Bcl-2相关X蛋白(Bcl-2-associated X protein, Bax)的表达;免疫组化法检测各组细胞和小鼠肿瘤组织中IL-6、磷酸化JAK2(phosphorylated JAK2, p-JAK2)、磷酸化STAT3(phosphorylated STAT3, p-STAT3)的表达。结果显示,lncRNA PVT1、 KLF12在宫颈癌组织和宫颈癌细胞系中的表达显著升高,miR-484的表达显著降低(P<0.05)。双荧光素酶试验证实,lncRNA PVT1靶向调控miR-484的表达;miR-484靶向调控KLF12的表达。抑制lncRNA PVT1能明显抑制宫颈癌细胞的增殖、迁移、侵袭,诱导细胞趋于凋亡,下调KLF12、 Bcl-2、 IL-6、 p-JAK2和p-STAT3的表达,上调Bax的表达(P<0.05),但antagomir能部分解除抑制lncRNA PVT1对宫颈癌进展的阻滞作用,而pcDNA3.1能部分恢复抑制lncRNAPVT1对宫颈癌进展的阻滞作用。该研究提示,在宫颈癌中lncRNA PVT1内源性竞争miR-484,上调KLF12的表达,促进宫颈癌细胞的体外增殖、迁移和侵袭,抑制细胞凋亡,并促进肿瘤细胞的体内生长与转移,推动宫颈癌的恶性进展,这可能与激活IL-6/STAT3信号通路有关。
The aim of this study is to investigate the effects of long non-coding RNA(lncRNA)plasmacytoma variant translocation 1(PVT1)on the proliferation,apoptosis,migration,and invasion of cervical cancer cells,and its regulation on miR-484 and Krüppel-like factor 12(KLF12),as well as the IL-6/STAT3 signaling pathway.qRT-PCR was used to detect the expressions of lncRNA PVT1,miR-484,and the mRNA expression of KLF12 in cervical cancer tissue and cell lines.Double luciferase assay was used to detect the relationship between lncRNA PVT1,miR-484,and KLF12.Upon transfection of Sh-PVT1,Sh-PVT1+antagomir,or Sh-PVT1+antagomir+pcDNA3.1,the proliferation,apoptosis,migration,and invasion of cells were examined.The in vivo tumor growth and metastasis were also measured.Western blotting was used to detect the expression of KLF12,B-cell lymphoma 2(Bcl-2),and Bcl-2-associated X protein(Bax)in each group of tumor cells and mouse tumor tissue.Immunohistochemistry was used to detect the expression of IL-6,phosphorylated JAK2(p-JAK2),and phosphorylated STAT3(p-STAT3)in cells and mouse tumor tissue.The results showed that the expressions of lncRNA PVT1 and KLF12 were significantly increased in cervical cancer tissue and cervical cancer cell lines,while the expression of miR-484 was significantly decreased(P<0.05).Double luciferase assay confirmed the targeted regulation of lncRNA PVT1 on the expression of miR-484 which in turn regulated the expression of KLF12.Inhibiting lncRNA PVT1 significantly inhibited the proliferation,migration,and invasion of cervical cancer cells,induced cell apoptosis,downregulated the expressions of KLF12,Bcl-2,IL-6,p-JAK2,and p-STAT3,and upregulated the expression of Bax(P<0.05).However,antagomir could partially relieve the suppression of inhibition of lncRNA PVT1 in cervical cancer progression,while pcDNA3.1 partially restored the suppression of inhibition of lncRNA PVT1 in cervical cancer progression.Therefore,this study suggests that in cervical cancer,lncRNA PVT1 upregulates KLF12 by endogenous competition with miR-484 to promote the proliferation,migration,and invasion of cervical cancer cells,inhibit apoptosis,promote the growth and metastasis of tumor cells,and the malignant progression of cervical cancer in vitro,which may be related to the activation of IL-6/STAT3 signaling pathway.
作者
包克勇
王东
包丽红
张莉
张雪
陶晓玉
高涵
BAO Ke-yong;WANG Dong;BAO Li-hong;ZHANG Li;ZHANG Xue;TAO Xiao-yu;GAO Han(Department of Gynecology,Affiliated Hospital of Inner Mongolia University for Nationalities,Tongliao 028000,China;Department of Oncology,Affiliated Hospital of Inner Mongolia University for Nationalities,Tongliao 028000,China;School of Medicine,Inner Mongolia University for Nationalities,Tongliao 028000,China)
出处
《现代免疫学》
2025年第2期151-163,共13页
Current Immunology
基金
内蒙古自治区自然科学基金(2022MS08077)。
作者简介
通信作者:包克勇(1976-),男,硕士,副主任医师,主要从事妇科肿瘤研究(E-mail:168163001@qq.com)。
