HHV-6A感染对神经胶质瘤细胞LncRNA MEG3表达及增殖凋亡侵袭转化的影响

doi:10.3969/j.issn.1004-616x.2019.05.003

癌变·畸变·突变 ›› 2019, Vol. 31 ›› Issue (5): 352-358.doi: 10.3969/j.issn.1004-616x.2019.05.003

HHV-6A感染对神经胶质瘤细胞LncRNA MEG3表达及增殖凋亡侵袭转化的影响

万昕¹, 胡月², 章菊¹, 陈云³, 金佩佩¹

1. 中国科学技术大学附属第一医院/安徽省立医院检验科, 安徽合肥 230036;
2. 中国科学技术大学附属第一医院/安徽省立医院妇产科产前诊断中心, 安徽合肥 230036;
3. 南京医科大学微生物与免疫学系, 江苏南京 211100

收稿日期:2019-03-15 修回日期:2019-06-19 出版日期:2019-09-30 发布日期:2019-10-09
通讯作者: 金佩佩,E-mail:kaixin@mail.ustc.edu.cn E-mail:kaixin@mail.ustc.edu.cn
作者简介:万昕,E-mail:wanxin242813@163.com。
基金资助:
国家自然科学基金青年科学基金项目（81701823）

Human herpesvirus 6A subtype affects the expression of LncRNA MEG3,proliferation,apoptosis and EMT process in glioma cells

WAN Xin¹, HU Yue², ZHANG Ju¹, CHEN Yun³, JIN Peipei¹

1. Department of Laboratory Medicine, The First Affiliated Hospital of University of Science and Technology of China/Anhui Provincial Hospital, Hefei 230036;
2. Prenatal Diagnosis Center, The First Affiliated Hospital of USTC/Anhui Provincial Hospital, Hefei 230036, Anhui;
3. Department of Microbiology and Immunology, Nanjing Medical University, Nanjing 211100, Jiangsu, China

Received:2019-03-15 Revised:2019-06-19 Online:2019-09-30 Published:2019-10-09

摘要/Abstract

摘要： 目的：选取胶质瘤相关长链非编码RNA（LncRNA）MEG3，观察人类疱疹病毒6A亚型（HHV-6A）感染对神经胶质瘤细胞LncRNA MEG3表达及其增殖、凋亡、侵袭、转化的影响。方法：实时定量PCR检测人神经胶质瘤细胞和正常神经胶质细胞，以及37例胶质瘤组织、4例癌旁组织、18例正常脑组织中LncRNA MEG3的表达。细胞计数试剂盒检测HHV-6A感染后神经胶质瘤细胞吸光度D（450）值，流式细胞术检测细胞凋亡率，划痕实验观察细胞划痕愈合度，蛋白质印迹法检测细胞上皮间质转化（EMT）指标蛋白的表达水平。结果：在胶质瘤组织或细胞中，LncRNA MEG3的表达水平较正常对照组织或细胞低（P < 0.05），而HHV-6A感染后的正常神经胶质细胞或胶质瘤细胞中LncRNA MEG3的表达也呈降低趋势（P < 0.05）。流式细胞术结果显示病毒感染后胶质瘤细胞的凋亡率为（6.05±0.40）%，对照组细胞凋亡率为（8.23±0.75）%，HHV-6A感染组凋亡率较对照组显著降低（P < 0.05）。划痕实验结果显示HHV-6A感染后细胞划痕愈合度[（72.33±6.90）%]大于对照组细胞划痕愈合度[（43.67±6.30）%]，差异有统计学意义（P < 0.05）。Western blot结果显示，与对照组相比，病毒感染后细胞中Snail及Vimentin蛋白表达水平升高，E-cadherin蛋白的表达降低（P < 0.05）。结论：HHV-6A感染可下调神经胶质细胞及胶质瘤细胞中LncRNA MEG3的表达，促进胶质瘤细胞增殖侵袭和上皮间质转化，并抑制其凋亡。

关键词: 神经胶质瘤, 长链非编码RNA MEG3, 人类疱疹病毒6A亚型, 上皮间质转化

Abstract: OBJECTIVE:The glioma-associated long non-coding RNA (LncRNA) MEG3 was preliminarily selected and detected in glioma tissues. Its expression in glioma cells together with proliferation,apoptosis,invasion and transformation of glioma cells were investigated after HHV-6A infection. METHODS:Expression of LncRNA MEG3 in 37 glioma,4 para-tumor tissues and 18 normal brain tissues,or HHV-6A infected glioma cells were measured by Real-time quantitative PCR. The D(450) value of HHV-6A infected glioma cells were assayed by using cell counting kit-8 method. The frequencies of apoptotic cells were detected by flow cytometry. The cell healing ability was monitored by the wound-healing assay. The epithelial-mesenchymal transition (EMT)-related proteins were evaluated by Western blot. RESULTS:Expression of LncRNA MEG3 was significantly lower in glioma tissues than in para-tumor tissues (P < 0.05). Expression of LncRNA MEG3 showed a down-regulation in glioma or normal glial cells after HHV-6A infection (P < 0.05). The apoptotic rates of HHV-6A infected glioma cells were significantly lower than that in the control group[(6.05±0.40)% and (8.23±0.75)%,respectively,P < 0.05]. The wound healing ability of glioma cells after HHV-6A infection was significantly better than that of the control group[(72.33±6.90)% and (43.67±6.30)%,respectively,P < 0.05]. Compared with the control group,expression of Snail and Vimentin were increased,while expression of Ecadherin was decreased in glioma cells after HHV-6A infection (P < 0.05). CONCLUSION:HHV-6A infection down-regulated the expression of LncRNA MEG3 in glial cells and glioma cells. The expression was associated with proliferation,invasion,epithelial-mesenchymal transition of glioma cells and inhibition of apoptosis.

Key words: glioma, long non-coding RNA MEG3, human herpesvirus 6A, epithelial-mesenchymal transition

中图分类号:

R730.23

万昕, 胡月, 章菊, 陈云, 金佩佩. HHV-6A感染对神经胶质瘤细胞LncRNA MEG3表达及增殖凋亡侵袭转化的影响[J]. 癌变·畸变·突变, 2019, 31(5): 352-358.

WAN Xin, HU Yue, ZHANG Ju, CHEN Yun, JIN Peipei. Human herpesvirus 6A subtype affects the expression of LncRNA MEG3,proliferation,apoptosis and EMT process in glioma cells[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2019, 31(5): 352-358.

参考文献

[1] MILLER J J, WEN P Y. Emerging targeted therapies for glioma[J]. Expert Opin Emerg Drugs, 2016, 21(4):441-452.
[2] REIFENBERGER G, WIRSCHING H G, KNOBBE-THOMSEN C B, et al. Advances in the molecular genetics of gliomas-implications for classification and therapy[J]. Nat Rev Clin Oncol, 2017, 14(7):434-452.
[3] CHEN R, SMITH-COHN M, COHEN A L, et al. Glioma subclassifications and their clinical significance[J]. Neurotherapeutics, 2017, 14(2):284-297.
[4] 陈宝师, 晋强, 张忠, 等. 联合靶向治疗复发恶性脑胶质瘤的研究[J]. 中华神经医学杂志, 2015, 14(7):740-742.
[5] DE BOLLE L, NAESENS L, DE CLERCQ E. Update on human herpesvirus 6 biology, clinical features, and therapy[J]. Clin Microbiol Rev, 2005, 18(1):217-245.
[6] CRAWFORD J R, SANTI M R, THORARINSDOTTIR H K, et al. Detection of human herpesvirus-6 variants in pediatric brain tumors:association of viral antigen in low grade gliomas[J]. J Clin Virol, 2009, 46(1):37-42.
[7] CHI J, GU B, ZHANG C, et al. Human herpesvirus 6 latent infection in patients with glioma[J]. J Infect Dis, 2012, 206(9):1394-1398.
[8] FAN Q, YANG L, ZHANG X D, et al. The emerging role of exosome-derived non-coding RNAs in cancer biology[J]. Cancer Lett, 2018, 414:107-115.
[9] WIECZOREK E, RESZKA E. mRNA, microRNA and lncRNA as novel bladder tumor markers[J]. Clin Chim Acta, 2018, 477:141-153.
[10] WANG L, YU Z T, SUN S Q, et al. Long non-coding RNAs:potential molecular biomarkers for gliomas diagnosis and prognosis[J]. Rev Neurosci, 2017, 28(4):375-380.
[11] HU C G, ZHOU Y F, LIU C, et al. Risk assessment model constructed by differentially expressed lncRNAs for the prognosis of glioma[J]. Oncol Rep, 2018, 40(5):2467-2476.
[12] DHAMIJA S, DIEDERICHS S. From junk to master regulators of invasion:lncRNA functions in migration, EMT and metastasis[J]. Int J Cancer, 2016, 139(2):269-280.
[13] ZHANG T, WANG Y R, ZENG F, et al. LncRNA H19 is overexpressed in glioma tissue, is negatively associated with patient survival, and promotes tumor growth through its derivative miR-675[J]. Eur Rev Med Pharmacol Sci, 2016, 20(23):4891-4897.
[14] QIN N, TONG G F, SUN L W, et al. Long noncoding RNA MEG3 suppresses glioma cell proliferation, migration, and invasion by acting as a competing endogenous RNA of miR-19a[J]. Oncol Res, 2017, 25(9):1471-1478.
[15] LIU F T, ZHU P Q, OU Y X, et al. Long non-coding RNA-LET can indicate metastasis and a poor prognosis:a meta-analysis[J]. Minerva Med, 2016, 107(2):101-107.
[16] MILLER G. Brain cancer. A viral link to glioblastoma?[J]. Science, 2009, 323(5910):30-31.
[17] CRAWFORD J R, SANTI M R, CORNELISON R, et al. Detection of human herpesvirus-6 in adult central nervous system tumors:predominance of early and late viral antigens in glial tumors[J]. J Neurooncol, 2009, 95(1):49-60.
[18] PARK J Y, LEE J E, PARK J B, et al. Roles of long non-coding RNAs on tumorigenesis and glioma development[J]. Brain Tumor Res Treat, 2014, 2(1):1-6.
[19] OMBRATO L, MALANCHI I. The EMT universe:space between cancer cell dissemination and metastasis initiation[J]. Crit Rev Oncog, 2014, 19(5):349-361.
[20] NIETO M A, HUANG R Y J, JACKSON R A, et al. Emt:2016[J]. Cell, 2016, 166(1):21-45.
[21] ZHANG L, LIANG X, LI Y X. Long non-coding RNA MEG3 inhibits cell growth of gliomas by targeting miR-93 and inactivating PI₃K/AKT pathway[J]. Oncol Rep, 2017, 38(4):2408-2416.

HHV-6A感染对神经胶质瘤细胞LncRNA MEG3表达及增殖凋亡侵袭转化的影响

Human herpesvirus 6A subtype affects the expression of LncRNA MEG3,proliferation,apoptosis and EMT process in glioma cells

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