碳水化合物磺基转移酶-11对肾透明细胞癌的生物学行为影响

Effect of carbohydrate sulfotransferase 11 on biological behavior in clear cell renal cell carcinoma

目的探讨碳水化合物磺基转移酶11(CHST11)对肾透明细胞癌(ccRCC)细胞增殖、迁移、侵袭生物学功能的影响。方法使用癌症基因组图谱(TCGA)数据库探究ccRCC组织与癌旁组织中CHST11基因的表达差异,分析CHST11表达水平与ccRCC分期、分级、预后的相关性;收集2021年1月至2021年10月在山西医科大学第一医院泌尿外科行手术切除的35例ccRCC和癌旁组织,采用荧光定量聚合酶链反应(RT-qPCR)、免疫组织化学和蛋白质印迹法(Western blot)检测ccRCC组织、癌旁组织及ccRCC细胞系中CHST11基因和蛋白表达水平;采用RNA干扰技术构建CHST11低表达的786O、769P细胞系;采用细胞计数试剂盒(CCK-8)实验方法检测细胞的增殖能力,采用Transwell实验方法检测细胞的迁移和侵袭能力。使用Graphpad Prism 9.0进行数据分析,Student’s t检验用于两组间比较,单因素方差分析用于多组间比较。结果生物信息学分析表明CHST11在ccRCC组织中mRNA表达水平高于癌旁组织(4.459±0.060比2.437±0.738,t=15.818,P<0.01),并且与患者预后生存率明显相关[风险比(HR)=1.490,P<0.01];实验结果表明,CHST11在ccRCC组织中,mRNA(1.865±1.705比1.000±0.956,t=4.949,P<0.01)和蛋白表达水平(1.989±0.353比1.000±0.539,t=4.949,P<0.01)高于癌旁组织;CHST11主要定位于细胞质与细胞膜,敲减CHST11基因可抑制ccRCC细胞增殖、迁移与侵袭的能力。结论CHST11在ccRCC中发挥促癌作用,沉默CHST11基因可抑制ccRCC细胞增殖、迁移与侵袭的能力。

Objective To investigate the impact of carbohydrate sulfotransferase 11(CHST11)on the biological functions of cell proliferation,migration,and invasion in clear cell renal cell carcinoma(ccRCC).Methods The differential expression of the CHST11 gene in ccRCC tissues and adjacent tissues was explored,and the correlation between CHST11 expression levels and ccRCC staging,grading,and prognosis value was analyzed using The Cancer Genome Atlas(TCGA)database.A total of 35 cases of ccRCC and adjacent tissues resected were selected during surgery in the Department of Urology,First Hospital of Shanxi Medical University from January 2021 to October 2021,and quantitative real time polymerase chain reaction(RT-qPCR),immunohistochemistry,and Western blotting were used to detect the gene and protein expression levels of CHST11 in ccRCC tissues,adjacent tissues,and ccRCC cell lines.The 786O and 769P cell lines with knockdown of CHST11 were constructed using RNA interference technology.The proliferative ability of cells was tested using the cell counting kit-8(CCK-8)assay,and the migration and invasion abilities of cells were measured using the Transwell assay.Data analysis was performed using Graphpad Prism 9.0,Student’s t-test was used for comparison between two groups,and one-way analysis of variance was used for comparison between multiple groups,with P<0.05 defining statistically significant difference.Results The bioinformatics analysis revealed that the mRNA expression level of CHST11 in ccRCC tissue was higher than that in the adjacent normal tissue(4.459±0.060 vs.2.437±0.738,t=15.818,P<0.01),and it was significantly correlated with patients’prognosis survival rate[hazard ratio(HR)=1.490,P<0.01].The experimental results indicated that in ccRCC tissue,the mRNA(1.865±1.705 vs.1.000±0.956,t=4.949,P<0.01)and protein expression levels(1.989±0.353 vs.1.000±0.539,t=4.949,P<0.01)of CHST11 were higher than in normal tissue.CHST11 was mainly located in the cytoplasm and cell membrane,and knocking down the CHST11 gene could inhibit the proliferation,migration,and invasion ability of ccRCC cells.Conclusion CHST11 plays a tumorigenic role in ccRCC,and silencing CHST11 gene can inhibit the proliferation,migration,and invasion of ccRCC cells.