miR-24-3p靶向CHD5影响甲状腺癌细胞迁移能力、侵袭能力和放射敏感性

doi:10.3969/j.issn.1673-8640.2023.07.005

摘要/Abstract

摘要：

目的探讨miR-24-3p调控染色体质域解螺旋酶DNA结合蛋白5（CHD5）表达对甲状腺癌细胞迁移能力、侵袭能力和放射敏感性的影响。方法选取2018年9月—2020年9月杭州市萧山区第一人民医院首次确诊为甲状腺癌的患者125例，收集其癌组织和癌旁组织（距肿瘤边缘>4 cm）和相关临床病理资料。选取人正常甲状腺细胞系Nthy-ori 3-1和人甲状腺癌细胞系KTC1、SW579、TPC-1。检测癌组织、癌旁组织和4种细胞中miR-24-3p、CHD5蛋白的表达。对SW579细胞分别转染miR-24-3p抑制物anti-miR-24-3p（anti-miR-24-3p组）和阴性对照物anti-miR-NC（anti-miR-NC组）、miR-24-3p模拟物miR-24-3p mimics（miR-24-3p mimics组）和阴性对照物miR-NC（miR-NC组）、共转染anti-miR-24-3p和si-NC（anti-miR-24-3p+si-NC组）、共转染anti-miR-24-3p和si-CHD5（anti-miR-24-3p+si-CHD5组）。分析各组细胞的迁移能力、增殖能力和放射敏感性。采用双荧光素酶报告基因实验验证miR-24-3p与CHD5的靶向关系。结果癌组织和甲状腺癌细胞系KTC1、SW579、TPC-1中miR-24-3p相对表达量均显著高于癌旁组织和正常甲状腺细胞系Nthy-ori 3-1（P<0.05），CHD5蛋白相对表达量均显著低于癌旁组织和正常甲状腺细胞系Nthy-ori 3-1（P<0.05）。SW579细胞miR-24-3p相对表达量最高，CHD5蛋白相对表达量最低。有无淋巴转移、不同临床分期的甲状腺癌患者之间miR-24-3p和CHD5蛋白表达差异有统计学意义（P<0.001）。抑制miR-24-3p表达可抑制SW579细胞的迁移能力和侵袭能力，并提高SW579细胞的放射敏感性（P<0.05）；miR-24-3p靶向负调控CHD5的表达；沉默CHD5可逆转抑制miR-24-3p表达对SW579细胞迁移、侵袭和放射敏感性的影响（P<0.05）。结论抑制miR-24-3p表达可上调CHD5，进而抑制SW579细胞的迁移能力和侵袭能力，提高其放射敏感性。

关键词: 微小RNA-24-3p, 染色体质域解螺旋酶DNA结合蛋白5, 放射敏感性, 细胞迁移, 细胞侵袭, 甲状腺癌

Abstract:

Objective To investigate the effects of microRNA（miR）-24-3p regulating the expression of chromodomain helicase DNA-binding protein 5（CHD5） on the migration，invasion and radiosensitivity of thyroid cancer cells. Methods A total of 125 patients with thyroid cancer were enrolled in Xiaoshan First People's Hospital from September 2018 to September 2020. Cancer tissues，paracancer tissues （4 cm from tumor margin） and clinicopathologic data were collected. Human normal thyroid cell line Nthy-ori 3-1 and human thyroid cancer cell lines，KTC1，SW579 and TPC-1，were selected. The expressions of miR-24-3p and CHD5 proteins in cancer tissue，paracancer tissue and 4 kinds of cells were determined. SW579 cells were transfected with miR-24-3p inhibitor anti-miR-24-3p（anti-miR-24-3p group），negative control anti-miR-NC （anti-miR-NC group），miR-24-3p mimic miR-24-3p mimics （miR-24-3p mimics group），negative control miR-NC （miR-NC group），co-transfected anti-miR-24-3p and si-NC （anti-miR-24-3p+si-NC group），co-transfected anti-miR-24-3p and si-CHD5 （anti-miR-24-3p+si-CHD5 group）. The migration ability，proliferation ability and radiosensitivity of each group were determined. Dual luciferase reporter assay was used to verify the targeting relationship between miR-24-3p and CHD5. Results The relative expression level of miR-24-3p in cancer tissue and thyroid cancer cell lines（KTC1，SW579 and TPC-1） were higher than those in paracancer tissue and normal thyroid cell line Nthy-ori 3-1（P<0.05）. The relative expression level of CHD5 protein was lower than those in paracancer tissue and normal thyroid cell line Nthy-ori 3-1（P<0.05）. SW579 cells had the highest relative expression of miR-24-3p and the lowest relative expression of CHD5 protein. The expressions of miR-24-3p and CHD5 proteins had statistical significance in thyroid cancer patients with or without lymphatic metastasis and different clinical stages（P<0.001）. The inhibition of miR-24-3p expression could inhibit the migration and invasion abilities of SW579 cells，and the radiosensitivity of SW579 cells was enhanced（P<0.05）. The targeting of miR-24-3p regulated CHD5 expression negatively. Silencing CHD5 could reverse the effects of miR-24-3p expression on migration，invasion and radiosensitivity of SW579 cells（P<0.05）. Conclusions The inhibition of miR-24-3p expression can inhibit the migration and invasion abilities of SW579 cells and enhance their radiosensitivity by up-regulating CHD5.

Key words: MicroRNA-24-3p, Chromodomain helicase DNA-binding protein 5, Radiosensitivity, Cell migration, Cell invasion, Thyroid cancer

中图分类号:

R393

潘信, 吕颖, 李阳, 李荣国. miR-24-3p靶向CHD5影响甲状腺癌细胞迁移能力、侵袭能力和放射敏感性[J]. 检验医学, 2023, 38(7): 640-646.

PAN Xin, LÜ Ying, LI Yang, LI Rongguo. Effects of miR-24-3p targeting CHD5 on migration，invasion and radiosensitivity of thyroid cancer cells[J]. Laboratory Medicine, 2023, 38(7): 640-646.

图/表 14

参考文献 20

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临床病例特征	例数	miR-24-3p	CHD5蛋白
性别
男性	40	2.27±0.31	0.18±0.05
女性	85	2.24±0.28	0.17±0.01
t值		0.540	0.915
P值		0.590	0.362
年龄
≤60岁	65	2.29±0.38	0.17±0.01
>60岁	60	2.21±0.19	0.17±0.03
t值		1.470	0.000
P值		0.144	1.000
淋巴转移
有	52	2.35±0.32	0.15±0.03
无	73	2.11±0.25	0.20±0.01
t值		4.704	13.261
P值		<0.001	<0.001
临床分期
Ⅰ期	78	2.05±0.27	0.14±0.04
Ⅱ~Ⅲ期	47	2.37±0.30	0.19±0.01
t值		6.154	10.532
P值		<0.001	<0.001

临床病例特征	例数	miR-24-3p	CHD5蛋白
性别
男性	40	2.27±0.31	0.18±0.05
女性	85	2.24±0.28	0.17±0.01
t值		0.540	0.915
P值		0.590	0.362
年龄
≤60岁	65	2.29±0.38	0.17±0.01
>60岁	60	2.21±0.19	0.17±0.03
t值		1.470	0.000
P值		0.144	1.000
淋巴转移
有	52	2.35±0.32	0.15±0.03
无	73	2.11±0.25	0.20±0.01
t值		4.704	13.261
P值		<0.001	<0.001
临床分期
Ⅰ期	78	2.05±0.27	0.14±0.04
Ⅱ~Ⅲ期	47	2.37±0.30	0.19±0.01
t值		6.154	10.532
P值		<0.001	<0.001

组别	miR-24-3p	迁移细胞数/（个/高倍镜视野）	侵袭细胞数/（个/高倍镜视野）
anti-miR-NC组	1.04±0.12	89.98±6.32	67.25±4.12
anti-miR-24-3p组	0.32±0.04	36.25±2.83	28.52±2.33
t值	13.943	19.006	20.043
P值	<0.001	<0.001	<0.001

组别	miR-24-3p	迁移细胞数/（个/高倍镜视野）	侵袭细胞数/（个/高倍镜视野）
anti-miR-NC组	1.04±0.12	89.98±6.32	67.25±4.12
anti-miR-24-3p组	0.32±0.04	36.25±2.83	28.52±2.33
t值	13.943	19.006	20.043
P值	<0.001	<0.001	<0.001

组别	存活率/%
组别	0 Gy	2 Gy	4 Gy	6 Gy	8 Gy
anti-miR-NC组	99.81±0.22	87.53±3.31	72.91±2.10	36.80±1.42	7.34±0.82
anti-miR-24-3p组	99.92±0.11	68.24±2.93	8.62±0.33	1.24±0.22	0.40±0.13
t值	1.095	10.689	74.080	60.618	20.475
P值	0.299	<0.001	<0.001	<0.001	<0.001