长链非编码RNA HCG22在口腔鳞状细胞癌中的表达及作用机制研究

doi:10.7518/hxkq.2021.06.006

华西口腔医学杂志 ›› 2021, Vol. 39 ›› Issue (6): 658-666.doi: 10.7518/hxkq.2021.06.006

长链非编码RNA HCG22在口腔鳞状细胞癌中的表达及作用机制研究

高永强¹(), 施鹏伟², 师文楷³, 刘一鸣²()

^1.郑州大学第一附属医院口腔牙周科，郑州 450052
^2.郑州大学第一附属医院口腔颌面外科，郑州 450052
^3.郑州大学第一附属医院肝胆外科，郑州 450052

收稿日期:2020-11-27 修回日期:2021-09-03 出版日期:2021-12-01 发布日期:2021-12-03
通讯作者: 刘一鸣 E-mail:Yongqiang86@outlook.com;liulei_yxk@163.com
作者简介:高永强，主治医师，硕士，E-mail：Yongqiang86@outlook.com
基金资助:
2020年度河南省高等学校重点科研项目(20B320019)

Expression and mechanism of long non-coding RNA HCG22 in oral squamous cell carcinoma

Gao Yongqiang¹(), Shi Pengwei², Shi Wenkai³, Liu Yiming²()

^1.Dept. of Periodontics, The First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, China
^2.Dept. of Oral and Maxillofacial Surgery, The First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, China
^3.Dept. of Hepatobiliary Surgery, The First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, China

Received:2020-11-27 Revised:2021-09-03 Online:2021-12-01 Published:2021-12-03
Contact: Liu Yiming E-mail:Yongqiang86@outlook.com;liulei_yxk@163.com
Supported by:
Institution of Higher Education Key Scientific Research Project of Henan Province in 2020(20B320019)

摘要/Abstract

摘要： 目的

探讨长链非编码RNA（lncRNA）HCG22在口腔鳞状细胞癌（OSCC）中的表达及作用机制。

方法

检测OSCC组织和对应癌旁组织、OSCC细胞和正常口腔角质细胞HOK中HCG22的水平；转染过表达载体质粒上调SCC-25和HSC-3细胞中HCG22的表达，采用甲基噻唑基四唑（MTT）法、流式细胞仪、Transwell小室检测细胞的增殖、凋亡，迁移和侵袭能力变化，Western blotting检测细胞上皮间质转化相关蛋白的表达；荧光定量聚合酶链反应（RT-qPCR）检测细胞中微小RNA-650（miR-650）表达水平；双荧光素酶报告基因实验检测HCG22和miR-650靶向关系。

结果

与癌旁组织比较，OSCC组织中HCG22表达明显降低（P<0.05），且HCG22低表达患者预后生存期显著低于高表达组（P<0.05）；与HOK细胞比较，SCC-25、HN13、HSC-3和CAL-27细胞中HCG22表达明显下调（P<0.05）；上调HCG22表达可抑制SCC-25和HSC-3细胞增殖、迁移、侵袭，诱导细胞凋亡，上调上皮间质转化蛋白上皮细胞钙黏蛋白（E-cadherin）和下调N-钙黏着蛋白（N-cadherin）、波形蛋白（Vimentin）蛋白表达（P<0.05）。miR-650模拟物可使转染HCG22野生型质粒细胞的荧光素酶活性降低（P<0.05），且上调HCG22表达后SCC-25和HSC-3细胞中miR-650表达下降（P<0.05）。

结论

HCG22在OSCC中低表达，上调其表达可抑制OSCC细胞的增殖、迁移、侵袭和上皮间质转化，其作用机制可能与对miR-650的靶向调控有关。

关键词: 口腔鳞状细胞癌, 长链非编码RNA HCG22, 细胞迁移, 细胞侵袭, 微小RNA-650

Abstract: Objective

To investigate the expression and mechanism of the long non-coding RNA (lncRNA) HCG22 in oral squamous cell carcinoma (OSCC).

Methods

HCG22 levels were detected in the OSCC and adjacent tissues, OSCC cells, and normal oral keratinocytes. HCG22 expression in SCC-25 and HSC-3 cells was upregulated by transfection of the overexpressing plasmi dvector. Methyl thiazolyl tetrazolium (MTT) assay, flow cytometry, and Transwell assay were employed to detect changes in cell proliferation, apoptosis, migration, and invasion ability, while Western blotting was used to detect the expression of epithelial-mesenchymal transformation-related proteins. The expression level of miR-650 in the cells was detected by real-time quantitative polymerase chain reaction (RT-qPCR), and dual-luciferase reporter gene assay was applied to assess the targeting relationship between HCG22 and miR-650.

Results

Compared with that in adjacent tissues, the expression of HCG22 significantly decreased in OSCC tissues (P<0.05). Moreover, the prognostic survival of patients in the low-HCG22 expression group was significantly lower than that in the high-expression group (P<0.05). Compared with that in HOK cells, the expression of HCG22 was significantly lower in SCC-25, HN13, HSC-3, and CAL-27 cells (P<0.05). Upregulation of HCG22 expression could inhibit the proliferation, migration, invasion, and apoptosis of SCC-25 and HSC-3 cells, upregulatethe expression of E-cadherin, and downregulate the expression of N-cadherin and vimentin (P<0.05). miR-650 mimics could reduce the luciferase activity of HCG22 wild-type plasmid cells (P<0.05), and the expression of miR-650 in SCC-25 and HSC-3 cells decreased after upregulation of HCG22 expression (P<0.05).

Conclusion

HCG22 is expressed at low levels in OSCC. Upregulation of the expression of this lncRNA can inhibit the proliferation, migration, invasion, and epithelial-mesenchymal transition of OSCC cells. The mechanism of action of HCG22 may be related to its targeted regulation of miR-650.

Key words: oral squamous cell carcinoma, long non-coding RNA HCG22, cell migration, cell invasion, microRNA-650

中图分类号:

Q 786

高永强, 施鹏伟, 师文楷, 刘一鸣. 长链非编码RNA HCG22在口腔鳞状细胞癌中的表达及作用机制研究[J]. 华西口腔医学杂志, 2021, 39(6): 658-666.

Gao Yongqiang, Shi Pengwei, Shi Wenkai, Liu Yiming. Expression and mechanism of long non-coding RNA HCG22 in oral squamous cell carcinoma[J]. West China Journal of Stomatology, 2021, 39(6): 658-666.

图/表 8

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资料		n	HCG22［n（%）］		χ²值	P值
资料		n	低表达组	高表达组	χ²值	P值
性别	男	36	20（55.56）	16（44.44）	0.846	0.358
	女	32	20（62.50）	12（37.50）
年龄/岁	>60	39	22（56.41）	17（43.59）	0.744	0.388
	≤60	29	18（62.07）	11（37.93）
肿瘤大小/cm	≥3	41	21（51.22）	20（48.78）	7.553	0.006
	<3	27	19（70.37）	8（29.63）
肿瘤部位	舌部	38	22（57.89）	16（42.11）	0.111	0.946
	颊部	20	12（60.00）	8（40.00）
	其他	10	6（60.00）	4（40.00）
TNM 分期	Ⅰ~Ⅱ期	40	26（65.00）	14（35.00）	4.604	0.032
	Ⅲ~Ⅳ期	28	14（50.00）	14（50.00）
分化程度	低分化	42	27（64.29）	15（35.71）	3.998	0.046
	中、高分化	26	13（50.00）	13（50.00）
淋巴结转移	无	47	30（63.83）	17（36.17）	5.195	0.023
	有	21	10（47.62）	11（52.38）

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长链非编码RNA HCG22在口腔鳞状细胞癌中的表达及作用机制研究

Expression and mechanism of long non-coding RNA HCG22 in oral squamous cell carcinoma

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