LncRNA HCP5调节miR-27b-3p/H2AFZ轴对口腔鳞状细胞癌增殖侵袭迁移的机制研究

doi:10.3969/j.issn.1006-6233.2023.01.004

摘要
图/表
参考文献
相关文章 (15)

全文: PDF (2301 KB) HTML (1 KB)
输出: BibTeX | EndNote (RIS)

摘要 目的: 探讨长链非编码RNA(LncRNA)人类白细胞抗原复合物P5(HCP5)通过调节微小RNA(miR)-27b-3p/H2A组蛋白家族成员Z(H2AFZ)轴对口腔鳞状细胞癌(OSCC)增殖、侵袭、迁移的影响。方法: 收集2021年2月至2022年2月在本院行手术治疗的48例OSCC患者癌组织及邻近癌旁组织、OSCC细胞系(Tca-811、SCC-9、SCC-25、HN4)及人正常口腔角质形成细胞(hNOK),检测LncRNA HCP5、miR-27b-3p、H2AFZ mRNA表达水平;取对数生长期Tca-811细胞,分为对照组、小干扰RNA阴性对照(si NC)组、干扰HCP5表达(si HCP5)组、si HCP5+抑制物阴性对照(inhibitor NC)组、si HCP5+miR-27b-3p抑制物(miR-27b-3p inhibitor)组。双荧光素酶验证LncRNA HCP5与miR-27b-3p、miR-27b-3p与H2AFZ的靶向关系;qRT-PCR检测LncRNA HCP5、miR-27b-3p、H2AFZ mRNA表达水平;流式细胞仪及MTT法检测Tca-811细胞凋亡及增殖变化;Transwell实验检测细胞侵袭、迁移变化;Western blot检测细胞周期蛋白D1(CyclinD1)、活化型半胱天冬酶3(cleaved caspase-3)、基质金属蛋白酶2(MMP-2)、H2AFZ蛋白表达。结果: 与hNOK细胞和癌旁组织比较,OSCC细胞和组织中LncRNA HCP5、H2AFZ mRNA表达升高,miR-27b-3p表达下降(P<0.05)。miR-27b-3p与LncRNA HCP5、H2AFZ均存在靶向关系。si HCP5组细胞凋亡率、cleaved caspase-3、miR-27b-3p表达较si NC组、对照组升高(P<0.05),OD570(24h、48h)、侵袭、迁移细胞数及CyclinD1、MMP-2、HCP5、H2AFZ表达下降(P<0.05);抑制miR-27b-3p表达逆转了干扰HCP5对Tca-811细胞恶性行为的抑制作用(P<0.05)。结论: 干扰LncRNA HCP5可通过靶向上调miR-27b-3p、抑制H2AFZ表达,进而抑制OSCC的增殖、侵袭、迁移。

	服务

	把本文推荐给朋友
	加入我的书架
	加入引用管理器
	E-mail Alert
	RSS
	作者相关文章

关键词 ： LncRNA HCP5, miR-27b-3p/H2AFZ轴, 增殖, 侵袭, 迁移, 口腔鳞状细胞癌

Abstract：Objective: To investigate the influences of long non-coding RNA (LncRNA) human leukocyte antigen complex P5 (HCP5) proliferation, invasion and migration of oral squamous cell carcinoma (OSCC) by regulating the microRNA (miR)-27b-3p/H2A histone family member Z (H2AFZ) axis. Methods: The cancer tissues and adjacent paracancerous tissues, OSCC cell lines (Tca-811, SCC-9, SCC-25, HN4) and human normal oral keratin-forming cells (hNOK) were collected from 48 OSCC patients treated surgically at our hospital from 2021 to 2022, and the LncRNA HCP5, miR-27b-3p, H2AFZ mRNA expression levels; logarithmic growth phase Tca-811 cells were divided into control group, small interfering RNA negative control (si NC) group, interfering HCP5 expression (si HCP5) group, si HCP5+ inhibitor negative control (inhibitor NC) group, si HCP5+ miR-27b-3p inhibitor (miR-27b-3p) group. Double luciferase was used to verify the targeting relationship between LncRNA HCP5 and miR-27b-3p, miR-27b-3p and H2AFZ; qRT-PCR to detect the expression levels of LncRNA HCP5, miR-27b-3p and H2AFZ mRNA; flow cytometry and MTT to detect apoptosis and proliferation changes of Tca-811 cells. Transwell assay was performed to detect changes in cell invasion and migration; Western blot to detect expression of cyclinD1, cleaved caspase-3, matrix metalloproteinase 2 and H2AFZ protein. Results: Compared with hNOK cells and adjacent tissues, the expressions of LncRNA HCP5 and H2AFZ mRNA in OSCC cells and tissues were increased, and the expression of miR-27b-3p was decreased (P<0.05). There was a targeting relationship between miR-27b-3p with LncRNA HCP5, H2AFZ. Compared with the si NC group and the control group, the apoptosis rate, cleaved caspase-3 and miR-27b-3p expressions in the si HCP5 group were higher than those in the si NC group and the control group (P<0.05), the OD570 (24 h, 48 h), the number of invasive and migrating cells and the expressions of CyclinD1, MMP-2, HCP5, H2AFZ were lower (P<0.05); inhibiting the expression of miR-27b-3p reversed the inhibitory effect of interfering with HCP5 on the malignant behavior of Tca-811 cells (P<0.05). Conclusion: Interfering with LncRNA HCP5 can inhibit the proliferation, invasion and migration of OSCC by targeting up-regulation of miR-27b-3p and inhibiting the expression of H2AFZ.

Key words： LncRNA HCP5 miR-27b-3p/H2AFZ axis Proliferation Invasion Migration Oral squamous cell carcinoma

基金资助:河北省2022年度医科科学研究课题,(编号:20221387)

通讯作者: 邸永宾

引用本文:

史晓晶, 周金阔, 张晋弘, 邸永宾, 姜磊. LncRNA HCP5调节miR-27b-3p/H2AFZ轴对口腔鳞状细胞癌增殖侵袭迁移的机制研究[J]. 河北医学, 2023, 29(1): 18-25.
SHI Xiaojing, ZHOU Jinkuo, ZHANG Jinhong, et al. The Mechanism of LncRNA HCP5 Regulating miR-27b-3p/H2AFZ Axis on the Proliferation Invasion and Migration of Oral Squamous Cell Carcinoma. HeBei Med, 2023, 29(1): 18-25.

链接本文:

http://www.hbyxzzs.cn/CN/10.3969/j.issn.1006-6233.2023.01.004 或 http://www.hbyxzzs.cn/CN/Y2023/V29/I1/18

[1] 桂心伟,姚小武,陈仕生,等.NF-κB在口腔鳞状细胞癌及癌旁组织中的表达意义[J].口腔疾病防治,2019,27(6):360-364.
[2] Liang L,Kang H,Jia J.HCP5 contributes to cisplatin resistance in gastric cancer through miR-128/HMGA2 axis[J].Cell Cycle,2021,20(11):1080-1090.
[3] Chen Y,Chen G,Zhang B,et al.miR-27b-3p suppresses cell proliferation,migration and invasion by targeting LIMK1 in colorectal cancer[J].Int Clin Exp Pathol,2017,10(9):9251-9261.
[4] Lagos-Quintana M,Rauhut R,Lendeckel W,et al.Identification of novel genes coding for small expressed RNAs[J].Science (New York,NY),2020,294(5543):853-858.
[5] Tang J,Fang X,Chen J,et al.Long non-coding RNA (lncRNA) in oral squamous cell carcinoma:biological function and clinical application[J].Cancers (Basel),2021,13(23):5944.
[6] Li X,Chen B,Huang A,et al.LncRNA HCP5 enhances the proliferation and migration of cervical cancer via miR-216a-5p/CDC42 axis[J].Cancer,2022,13(6):1882-1894.
[7] Zhao J,Bai X,Feng C,et al.Long non-coding RNA HCP5 facilitates cell Invasion and epithelial-mesenchymal transition in oral squamous cell carcinoma by miR-140-5p/SOX4 axis[J].Cancer Manag Res,2019,11:10455-10462.
[8] Lu M,Liu D,Li Y.Long intergenic non-protein coding RNA 467 inhibition elevates microRNA-27b-3p to repress malignant behaviors of gastric cancer cells via reducing STAT3[J].Cell Death Discov,2022,8(1):100.
[9] Zhao L,Han S,Hou J,et al.The local anesthetic ropivacaine suppresses progression of breast cancer by regulating miR-27b-3p/YAP axis[J].Aging (Albany NY),2021,13(12):16341-16352.
[10] Ma SQ,Wang YC,Li Y,et al.LncRNA XIST promotes proliferation and cisplatin resistance of oral squamous cell carcinoma by downregulating miR-27b-3p[J].Biol Regul Homeost Agents,2020,34(6):1993-2001.
[11] Feng J,Li Y,Wen N.Characterization of cancer stem cell characteristics and development of a prognostic stemness index cell-related signature in oral squamous cell carcinoma[J].Dis Markers,2021,2021:1571421.