口腔鳞癌环状RNA差异表达谱分析及circ_PVT1致癌机制的研究

doi:10.13591/j.cnki.kqyx.2025.09.003

摘要/Abstract

摘要：

目的筛选口腔鳞状细胞癌(oral squamous cell carcinoma,OSCC)环状RNA(circular RNA,circ_RNA)差异表达谱,阐明circ_PVT1对OSCC致癌的分子机制。方法利用circ_RNA表达谱芯片,对3例OSCC及对照正常组织的转录本进行高通量测序,筛选差异基因表达谱,进行GO和KEGG富集分析。利用qRT-PCR检测OSCC组织、正常人口腔角质上皮细胞及OSCC细胞中PVT1的表达水平,借助MTT实验、Transwell实验及流式细胞术评估PVT1对SCC-25、SCC-9细胞生物学行为的影响,采用Western blot法评估PVT1对Wnt3a/β-catenin通路中关键蛋白表达的影响,进一步研究PVT1的表达与患者临床病理特征及预后的关系。结果芯片共筛选出差异表达circ_RNAs 403条,差异表达基因都在与癌症进展相关的通路中富集。PVT1在OSCC组织和细胞中高表达,沉默PVT1表达可抑制Wnt3a/β-catenin通路的激活从而有效抑制SCC-25、SCC-9细胞的增殖、迁移、侵袭和细胞周期,促进凋亡。PVT1表达只与患者淋巴结转移、远端转移相关且高表达患者有着较短的PFS。结论 PVT1通过调控Wnt3a/β-catenin通路的激活促进OSCC的发病进展,研究成果为OSCC的诊疗提供了新思路。

关键词: 口腔鳞癌, 基因芯片, circ_PVT1, Wnt3a/β-catenin, 通路

Abstract:

Objective To screen the differential expression profile of circ_RNA in OSCC and to elucidate the molecular mechanism of circ_PVT1 on OSCC carcinogenesis. Methods The transcripts of 3 cases of OSCC and normal tissues were sequenced by high-throughput sequencing using circ_RNA expression profile chip, and the differential gene expression profiles were screened, and GO and KEGG enrichment analysis were performed. The expression level of PVT1 in OSCC tissues, human normal oral mucosal cells and OSCC cells was detected by qRT-PCR. The effect of PVT1 on the biological behavior of SCC-25 and SCC-9 cells was evaluated by MTT experiment, Transwell experiment and flow cytometry. The effect of PVT1 on the expression of key proteins in the Wnt3a/β-catenin pathway was evaluated by Western blot. The relationship between the expression of PVT1 and clinical pathological characteristics and prognosis of patients was further studied. Results A total of 403 differentially expressed circ_RNAs were screened by the chip, and the differentially expressed genes were enriched in pathways related to cancer progression. PVT1 was highly expressed in OSCC tissues and cells. Silencing PVT1 expression could inhibit the activation of the Wnt3a/β-catenin pathway, thereby effectively inhibiting the proliferation, migration, invasion and cell cycle of SCC-25 and SCC-9 cells and promoting apoptosis. PVT1 expression was only associated with lymph node metastasis and distant metastasis in patients, and those with high expression had a shorter PFS. Conclusion PVT1 promotes the progression of OSCC by regulating the activation of Wnt3a/β-catenin pathway. The research results provide new ideas for the diagnosis and treatment of OSCC.

Key words: oral squamous cell carcinoma, gene chip, circ_PVT1, Wnt3a/β-catenin, pathway

中图分类号:

R739.85

吴丽玮, 仇永乐, 赵佳红, 庄志征, 李昆珊. 口腔鳞癌环状RNA差异表达谱分析及circ_PVT1致癌机制的研究[J]. 口腔医学, 2025, 45(9): 655-662.

WU Liwei, QIU Yongle, ZHAO Jiahong, ZHUANG Zhizheng, LI Kunshan. Analysis of differential expression profiles of circular RNA in oral squamous cell carcinoma and study on the carcinogenic mechanism of circ_PVT1[J]. Stomatology, 2025, 45(9): 655-662.

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siRNAs	上游引物	下游引物
PVT1-siRNA1	5'-GGCUUGAGGCCUGAUCUUUTT-3'	5'-AAAGAUCAGGCCUCAAGCCTT-3'
PVT1-siRNA2	5'-GGCUUGAGGCCUGAUCUUUTT-3'	5'-AAAGAUCAGGCCUCAAGCCTT-3'
PVT1-siRNA3	5'-GCUGGGCUUGAGGCCUGAUTT-3'	5'-AUCAGGCCUCAAGCCCAGCTT-3'
NC-siRNA	5'-GCUACGAUCUGCCUAAGAUTT-3'	5'-AUCUUAGGCAGAUCGUCGCTT-3'

circRNAs	Fold-change	P	Hostgene
上调
hsa_circ_0090990	33.273 63	0.000 58	ZMYM3
hsa_circ_0000423	32.632 89	0.002 32	PPP1R12A
hsa_circ_0036540	25.440 78	0.008 44	MESDC2
hsa_circ_0078337	21.192 85	0.005 08	IPCEF1
hsa_circ_0030556	12.168 26	0.007 03	MIR17HG
hsa_circ_0013128	7.115 96	0.008 25	CDC7
hsa_circ_0054291	6.853 83	0.004 89	ZFP36L2
hsa_circ_0070429	6.847 98	0.004 47	HERC3
hsa_circ_0009077	5.820 61	0.002 65	TYW1B
hsa_circ_0033689	5.770 88	0.000 25	ADAM6
下调
hsa_circ_0058498	43.273 63	0.008 62	COL4A4
hsa_circ_0008269	32.632 89	0.002 49	USP8
hsa_circ_0056924	22.440 78	0.007 76	SCN9A
hsa_circ_0008304	15.192 85	0.008 66	TFDP2
hsa_circ_0001323	10.168 26	0.001 16	BBX
hsa_circ_0038262	10.115 96	0.000 86	SMG1
hsa_circ_0068421	8.853 83	0.009 88	TRA2B
hsa_circ_0073527	6.847 98	0.009 35	FER
hsa_circ_0035196	5.820 61	0.002 36	ATP8B4
hsa_circ_0031817	5.770 88	0.009 87	METTL21D