hsa_circRNA_0000596促进宫颈癌细胞侵袭和转移的机制研究OACSTPCD
Mechanism of hsa_circRNA_0000596 in promoting invasion and metastasis of cervical cancer cells
目的 探讨hsa_circRNA_000596(circ-596)促进宫颈癌细胞侵袭和转移的作用机制.方法 收集69例宫颈鳞状细胞癌(CSCC)手术患者的CSCC组织、癌旁正常宫颈组织及临床资料.采用RNA原位杂交(RISH)技术检测circ-596阳性表达情况,并分析其与CSCC患者临床特征的关系.采用Transwell实验检测宫颈癌细胞的侵袭能力;采用实时荧光定量聚合酶链反应(qRT-PCR)实验检测circ-596对下游靶基因的影响;采用双荧光素酶报告基因实验验证miR-197-3p对下游靶基因RNF146的调控作用;采用Rescue实验分析circ-596对miR-197-3p/RNF146信号通路及细胞侵袭能力的影响.结果 circ-596在CSCC组织中主要呈阳性表达,阳性表达率为63.77％(44/69),高于癌旁正常组织的17.39％(12/69),差异有统计学意义(x2=8.254,P=0.004).circ-596阳性表达与肿瘤分化程度、淋巴结转移均密切相关(P＜0.05).Transwell实验结果显示,与对照细胞相比,过表达circ-596的宫颈癌细胞侵袭数目增加,敲低circ-596表达的细胞侵袭数目减少,差异均有统计学意义(P＜0.05).生物信息学预测结果显示,miR-197-3p是circ-596的下游靶基因,RNF146是miR-197-3p的下游靶基因.qRT-PCR实验结果显示,过表达circ-596能下调miR-197-3p的表达,降低circ-596表达能上调miR-197-3p的表达,差异有统计学意义(P＜0.05).双荧光素酶报告基因实验结果显示,miR-197-3p与RNF146基因的3'UTR结合,从而抑制荧光素酶相对活性.Rescue实验结果显示,circ-596能够抑制miR-197-3p的表达,进而促进RNF146的表达,提升宫颈癌细胞的侵袭能力.结论 CSCC组织中circ-596主要呈阳性表达,且其阳性表达情况与肿瘤分化程度和淋巴结转移密切相关.circ-596通过结合miR-197-3p,促进RNF146的表达,进而促进宫颈癌细胞的侵袭和转移.
Objective To investigate the mechanism of hsa_circRNA_000596(circ-596)in pro-moting invasion and metastasis of cervical cancer cells.Methods The cervical squamous cell carcino-ma(CSCC)tissue,normal cervical tissue adjacent to cancer and clinical data of 69 cases with CSCC were collected.RNA in situ hybridization(RISH)was used to detect the expression of circ-596 and ana-lyze its association with clinical data of CSCC.The invasive ability of cervical cancer cells was detected by Transwell assay;the regulation of downstream target gene by circ-596 was detected by quantitative re-verse transcriptase PCR(qRT-PCR).Dual luciferase reporting experiment was used to verify the regula-tion of miR-197-3p on the downstream target gene RNF146;the regulation of miR-197-3p/RNF146 by circ-596 and its effect on invasion ability were analyzed in rescue experiment.Results Circ-596 was mainly positively expressed in CSCC tissues,and the positive expression rate was 63.77％(44/69),which was higher than 17.39％(12/69)in adjacent normal tissues(x2=8.254,P=0.004).Circ-596 positive expression was closely correlated with tumor differentiation and lymph node metastasis(P＜0.05).The results of Transwell experiment showed that compared with the control cells,the number of cervical cancer cells with overexpression of circ-596 increased,and the number of cells with knockdown expression of circ-596 decreased(P＜0.05).Bioinformatics prediction results showed that miR-197-3p was the downstream target gene of circ-596,and RNF146 was the downstream tar-get gene of miR-197-3p.The results of qRT-PCR showed that overexpression of circ-596 could down-regulate the expression of miR-197-3p,and down-expression of circ-596 could up-regulate the expression of miR-197-3p(P＜0.05).The results of dual luciferase reporter gene experiments showed that miR-197-3p bound to the 3'UTR of RNF146 gene,thereby inhibiting the relative activi-ty of luciferase.Rescue experiment results showed that circ-596 could inhibit the expression of miR-197-3p,and thereby promoting the expression of RNF146,and enhancing the invasion ability of cer-vical cancer cells.Conclusion Circ-596 is mainly positively expressed in CSCC tissues,and its positive expression is closely related to the degree of tumor differentiation and lymph node metasta-sis.Circ-596 promotes the expression of RNF146 by binding to miR-197-3p,thereby promoting the invasion and metastasis of cervical cancer cells.
张广凤;陈珏;王成海
扬州大学医学院病理学教研室,江苏扬州,225009||江苏省宝应县人民医院妇科,江苏宝应,225800扬州大学附属医院肿瘤科,江苏扬州,225012扬州大学医学院病理学教研室,江苏扬州,225009
临床医学
宫颈鳞状细胞癌hsa_circRNA_0000596微小RNA-197-3pRNF146基因侵袭转移
cervical squamous cell carcinomahsa_circRNA_0000596microRNA 197-3pRNF146 geneinvasionmetastasis
《实用临床医药杂志》 2024 (020)
6-11,22 / 7
江苏省中医药科技发展计划项目(ZD202330)
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