LncRNA SNHG20靶向调控miR-520c-3p/RAB22A通路对人口腔鳞状细胞癌细胞上皮间质转化及微管形成的影响

doi:10.19723/j.issn.1671-167X.2025.01.005

摘要/Abstract

摘要：

目的: 探究LncRNA SNHG20靶向调控miR-520c-3p/RAB22A通路对人口腔鳞状细胞癌(oral squamous cell carcinoma，OSCC)细胞上皮间质转化(epithelial-mesenchymal transition，EMT)及微管形成的影响。方法: 检测OSCC细胞及组织中LncRNA SNHG20、miR-520c-3p、RAB22A mRNA水平及其相互间关系。将OSCC细胞分为对照组、sh-NC组、sh-SNHG20组、sh-SNHG20+anti-NC组、sh-SNHG20+anti-miR-520c-3p组, 检测OSCC细胞EMT蛋白表达，检测微管形成数量变化，裸鼠成瘤实验检测LncRNA SNHG20对OSCC肿瘤生长的影响。结果: OSCC组织和细胞中LncRNA SNHG20、RAB22A mRNA上调表达，miR-520c-3p下调表达(P < 0.05)；LncRNA SNHG20与miR-520c-3p、RAB22A与miR-520c-3p之间均有结合位点；与sh-NC组相比，sh-SNHG20组间质样细胞数量较少，上皮样细胞数量较多，微管结构不完整且结节数量较少，LncRNA SNHG20、RAB22A、N-cadherin、vimentin下调表达，miR-520c-3p、E-cadherin上调表达(P < 0.05)；与sh-SNHG20+anti-NC组相比，sh-SNHG20+anti-miR-520c-3p组间质样细胞数量较多，上皮样细胞数量较少，微管排列较紧密，微管结节数量较多，miR-520c-3p、E-cadherin下调表达，RAB22A、N-cadherin、vimentin上调表达(P < 0.05)。sh-SNHG20组比sh-NC组OSCC移植瘤体积较小，质量较低，LncRNA SNHG20、RAB22A下调表达，miR-520c-3p上调表达(P < 0.05)。结论: 抑制LncRNA SNHG20表达能够靶向调节miR-520c-3p/RAB22A通路抑制OSCC细胞EMT和微管形成。

关键词: 口腔鳞状细胞癌, 小核仁RNA宿主基因20, 微小RNA-520c-3p, Rab蛋白22a, 上皮间质转化, 微管形成

Abstract:

Objective: To investigate the effects of LncRNA SNHG20 on epithelial mesenchymal transition (EMT) and microtubule formation in human oral squamous cell carcinoma (OSCC) cells through targeted regulation of the miR-520c-3p/RAB22A pathway. Methods: After real-time fluorescence quantitative detection of LncRNA SNHG20, miR-520c-3p, RAB22A mRNA expression levels in OSCC tissues and cells, dual luciferase reporter assay was used to detect the relationship between the three. OSCC cells were randomly separated into control group, sh-NC group, sh-SNHG20 group, sh-SNHG20+anti NC group, and sh-SNHG20+anti miR-520c-3p group. Western blotting was used to detect the expression of N-cadherin, vimentin, and E-cadherin proteins in the OSCC cells. The morphology of HSC-3 cells was observed under microscope. Changes in the number of microtubules formed were detected. The effect of LncRNA SNHG20 on the growth of OSCC tumors and the expression levels of LncRNA SNHG20, miR-520c-3p and RAB22 A in the transplanted tumors were detected by nude mice tumorigenesis experiment. Results: LncRNA SNHG20 and RAB22A mRNA were upregulated in the OSCC tissues and cells, while miR-520c-3p was downregulated (P < 0.05). There were binding sites between LncRNA SNHG20 and miR-520c-3p, RAB22A and miR-520c-3p, which had targeted regulation relationship. Compared with the sh-NC group, the sh-SNHG20 group had fewer stromal like cells, more epithelial like cells, incomplete microtubule structure, and fewer nodules. LncRNA SNHG20, RAB22A, N-Cadherin, and vimentin were downregulated, while miR-520c-3p and E-cadherin were upregulated (P < 0.05). Compared with the sh-SNHG20+anti-NC group, the sh-SNHG20+anti-miR-520c-3p group had a higher number of stromal like cells, a lower number of epithelioid cells, tighter microtubule arrangement, and more microtubule nodules. miR-520c-3p and E-cadherin were downregulated, while RAB22A, N-cadherin, and vimentin were upregulated (P < 0.05). The transplanted tumor of OSCC in sh-SNHG20 group was smaller and lower than that in sh-NC group. The expression levels of LncRNA SNHG20 and RAB22A in the transplanted tumor tissues were lower than those in sh-NC group, and the expression level of miR-520c-3p was higher than that in sh-NC group (P < 0.05). Conclusion: LncRNA SNHG20 promotes epithelial-mesenchymal transition and microtubule formation in human oral squamous cell carcinoma cells by targeting the miR-520c-3p/RAB22A pathway. Inhibiting the expression of LncRNA SNHG20 can target and regulate the miR-520c-3p/RAB22A pathway to inhibit EMT and microtubule formation in OSCC cells.

Key words: Oral squamous cell carcinoma, Small nucleolar RNA host gene 20, Micro RNA-520c-3p, Rab protein 22a, Epithelial mesenchymal transition, Microtubule formation

中图分类号:

R739.8

马民英, 晁晓芹, 赵扬, 赵国廷. LncRNA SNHG20靶向调控miR-520c-3p/RAB22A通路对人口腔鳞状细胞癌细胞上皮间质转化及微管形成的影响[J]. 北京大学学报（医学版）, 2025, 57(1): 26-32.

Minying MA, Xiaoqin CHAO, Yang ZHAO, Guoting ZHAO. Effects of LncRNA SNHG20 on epithelial mesenchymal transition and microtubule formation in human oral squamous cell carcinoma cells through targeted regulation of the miR-520c-3p/RAB22A pathway[J]. Journal of Peking University (Health Sciences), 2025, 57(1): 26-32.

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Group	LncRNA SNHG20/GAPDH	miR-520c-3p/U6	RAB22A mRNA/GAPDH
TAC	0.57±0.05	1.13±0.11	0.52±0.05
OSCC	1.20±0.12	0.64±0.06	1.08±0.10
t	39.366	35.166	43.536
P	< 0.001	< 0.001	< 0.001

Group	LncRNA SNHG20/GAPDH	miR-520c-3p/U6	RAB22A mRNA/GAPDH
HIOEC	0.62±0.06	1.02±0.11	0.46±0.05
HSC-3	1.18±0.11	0.51±0.05	1.07±0.10
t	10.947	10.339	13.364
P	< 0.001	< 0.001	< 0.001

Group	LncRNA SNHG20/GAPDH	miR-520c-3p/U6	RAB22A protein/β-actin
Control	1.02±0.10	0.84±0.08	1.27±0.12
sh-NC	1.00±0.09	0.82±0.08	1.25±0.11
sh-SNHG20	0.52±0.05^a	1.29±0.12^a	0.58±0.05^a
sh-SNHG20+anti-NC	0.54±0.05	1.30±0.13	0.57±0.05
sh-SNHG20+anti-miR-520c-3p	0.53±0.05	0.95±0.09^b	0.80±0.08^b
F	81.117	32.557	95.090
P	< 0.001	< 0.001	< 0.001

Group	E-cadherin/β-actin	N-cadherin/β-actin	Vimentin/β-actin
Control	0.43±0.04	1.03±0.10	1.19±0.11
sh-NC	0.41±0.04	1.04±0.11	1.16±0.12
sh-SNHG20	0.93±0.09^a	0.65±0.06^a	0.70±0.07^a
sh-SNHG20+anti-NC	0.90±0.09	0.63±0.06	0.72±0.07
sh-SNHG20+anti-miR-520c-3p	0.58±0.05^b	0.87±0.08^b	0.95±0.09^b
F	86.233	33.008	36.642
P	< 0.001	< 0.001	< 0.001

Group	Tumor mass/g	Tumor volume/mm³	LncRNA SNHG20/GAPDH	miR-520c-3p/U6	RAB22A positive rate/%
sh-NC	1.35±0.13	1 476.98±156.39	1.17±0.11	1.00±0.11	47.45±5.08
sh-SNHG20	0.46±0.04	583.41±60.27	0.52±0.05	1.41±0.15	18.69±1.97
t	16.028	13.059	13.177	5.399	12.929
P	< 0.001	< 0.001	< 0.001	< 0.001	< 0.001