SHCBP1基因表达及对宫颈癌细胞生物学行为的影响研究

Effect of SHCBP1 gene expression on biological behavior of cervical cancer cells

目的 探讨宫颈上皮脱落细胞中SH2结构域结合蛋白1(SHCBP1)基因表达水平与宫颈病变程度、人乳头瘤病毒(HPV)感染及宫颈癌进展的相关性。方法 收集2022年7月—2023年10月343例患者的宫颈上皮脱落细胞,根据液基薄层细胞检测结果进行分组,其中121例非典型鳞状细胞,108例低度鳞状上皮内病变,56例高度鳞状上皮内病变,58例宫颈癌。采用Trizol法提取总RNA,采用RT-PCR法分析SHCBP1表达水平,统计分析SHCBP1表达水平与宫颈癌变的相关性。根据HPV结果,分析SHCBP1表达水平与HPV感染的相关性。根据UALCAN数据库、GEPIA数据库及临床组织样本,分析SHCBP1在宫颈癌及癌旁组织中的表达水平,利用慢病毒转染技术敲除宫颈癌细胞系SiHa中的SHCBP1表达后,利用MTT、划痕实验、克隆形成实验、Transwell实验及粘附实验分析SHCBP1表达水平对宫颈癌细胞增殖、迁移、粘附及克隆形成能力的影响,应用流式细胞仪分析SHCBP1表达对宫颈癌细胞周期阻滞的影响。结果 343例宫颈上皮脱落细胞中均有SHCBP1基因表达。非典型鳞状细胞、低度鳞状上皮内病变、高度鳞状上皮内病变、宫颈癌中SHCBP1表达水平分别为(3.75±1.49)、(4.10±1.56)、(7.41±1.51)、(11.52±2.13),差异有统计学意义(F=352.50,P<0.05)。SHCBP1基因表达水平与HPV感染相关,HPV16/HPV18阳性患者中SHCBP1表达最高。宫颈癌组织中SHCBP1基因表达水平最高。宫颈癌细胞SHCBP1表达水平与宫颈癌细胞恶性生物学行为密切相关。结论 宫颈上皮脱落细胞中SHCBP1表达水平与宫颈病变程度及HPV感染水平有关,其参与宫颈癌细胞恶性行为的生物学调控作用及其在癌及癌旁组织中的差异表达使SHCBP1有望成为宫颈病变进展及预测预后的新型标志物。

Objective To explore the correlations between SH_(2) domain-binding protein 1(SHCBP1)expression in cervical exfoliated cells and the degree of cervical lesions,human papillomavirus(HPV)infection,and cervical cancer progression.Methods The cervical epithelial exfoliated cells of 343 patients were collected and grouped according to the results of liquid-based thin layer cells from July 2022 to October 2023,including 121 cases of atypical squamous cells,108 cases of low-grade squamous intraepithelial lesion,56 cases of high-grade squamous intraepithelial lesion,and 58 cases of cervical cancer.Total RNA was extracted by Trizol method,SHCBP1 expression levels were analyzed by RT-PCR,and the correlation between SHCBP1 expression level and cervical carcinogenesis was statistically ana-lyzed.According to HPV results,the correlation between expression level of SHCBP1 and HPV infection was analyzed.The expression levels of SHCBP1 in cervical cancer and adjacent tissues were analyzed based on UALCAN database,GEPIA database,and clinical tissue sam-ples.After knocking out the expression of SHCBP1 in cervical cancer cell line SiHa using lentiviral transfection technology,the effect of SHCBP1 expression on proliferation,migration,adhesion,and clone formation abilities of cancer cell SiHa cells were analyzed using MTT assay,scratch test,clone formation test,Transwell assay,and adhesion experiment.The impact of SHCBP1 expression on cell cycle arrest of cancer cell SiHa cells was analyzed using flow cytometry.Results SHCBP1 gene expression in 343 cervical epithelial exfoliated cells were all found,and the expression levels in cases of atypical squamous cells,low-grade squamous intraepithelial lesion,high-grade squamous in-traepithelial lesion,and cervical cancer were(3.75±1.49),(4.10±1.56),(7.41±1.51),and(11.52±2.13),respectively,there was statistically significant difference(F=352.50,P<0.05).The expression level of SHCBP1 gene was associated with HPV infection,and the expression of SHCBP1 was the highest in HPV16+/HPV18+patients.The expression level of SHCBP1 in cervical cancer tissues was the highest.The expression level of SHCBP1 gene in cervical cancer cells was closely related to malignant biological behaviors of cervical cancer cells.Conclusion The expression level of SHCBP1 in cervical epithelial exfoliated cells is closely related to the degree of cervical lesion and HPV infection.The involvement in biological regulation of malignant behavior of cervical cancer cells and differential expression in cervi-cal cancer and adjacent tissues make SHCBP1 a promising new biomarker for cervical disease progression and prognosis prediction.