特异性蛋白1对肿瘤细胞增殖和转移的影响及其机制

Effect of transcription factor SP1 on proliferation and metastasis of cancer cells and underlying mechanism

目的:探讨特异性蛋白1(Sp1)对肿瘤细胞增殖和转移的影响及其机制。方法:选取2019年6月至2022年6月郑州大学第一附属医院收治的74例卵巢癌组织和癌旁组织作为研究对象,采用蛋白质印迹法(Western blot)分析癌旁组织和肿瘤组织的表达水平。采用SP1短发卡RNA(shRNA)慢病毒感染SK-OV-3细胞,建立SP KD1组和对照组细胞,采用细胞计数试剂盒(CCK-8)和EdU染色分析两组细胞的增殖能力;采用划痕实验和Transwell实验分析两组细胞的迁移和侵袭能力,采用蛋白质免疫印迹分析两组细胞上皮间质转化水平。组间资料比较采用t检验。结果:癌旁组织中SP1蛋白表达水平(1.03±0.22)明显低于卵巢癌组织(1.93±0.39),差异有统计学意义(t=17.570,P<0.05)。对照组细胞吸光度(A)值(1.98±0.09)明显高于SP KD1组(1.27±0.14),差异有统计学意义(t=10.310,P<0.05)。对照组细胞EdU阳性率[(84.17±4.07)%]明显高于SP KD1组[(65.67±5.57)%],差异有统计学意义(t=6.566,P<0.05)。对照组细胞划痕愈合率[(86.46±3.94)%]明显高于SP KD1组[(74.25±5.07)%],差异有统计学意义(t=4.659,P<0.05)。对照组细胞侵袭个数[139.67±12.09)个]明显高于SP KD1组[(97.67±9.52)个],差异有统计学意义(t=6.684,P<0.05)。对照组细胞间质细胞标志物N-钙黏蛋白(N-cadherin)和波形蛋白(Vimentin)(0.88±0.08、1.12±0.09)明显高于SP KD1组(0.68±0.08、0.78±0.07),差异有统计学意义(t=4.512、7.335,P<0.05)。对照组细胞上皮细胞标志物E-钙黏蛋白(E-cadherin)(0.81±0.06)明显低于SP KD1组(1.25±0.14),差异有统计学意义(t=7.156,P<0.05)。对照组细胞微小RNA(miR)-182表达水平(1.10±0.15)明显高于SP KD1组(0.79±0.08),差异有统计学意义(t=4.503,P<0.05)。结论:转录因子SP1在卵巢癌组织中呈高表达,通过提高miR-182表达水平,进而促进卵巢癌细胞的增殖和迁移能力。

Objective To investigate the effect of transcription factor SP1 on the proliferation and metastasis of cancer cells and the underlying mechanism.Methods Totally,74 cases of ovarian cancer and adjacent tissues admitted to first affiliated hospital of zhengzhou university from June 2019 to June 2022 were selected as the research subjects.The expression levels of adjacent tissues and tumor tissues were analyzed by Western blotting.SK-OV-3 cells were infected with SP1 short hairpin RNA(shRNA)Lentivirus to establish SPKD1 group and control group.The proliferation ability of the two groups was analyzed by cell counting kit-8(CCK-8)and EdU staining.The migration and invasion abilities of the two groups were analyzed using wounding healing and Transwell experiments and the levels of epithelial mesenchymal transformation were analyzed using Western blotting.The comparison of data between groups was conducted using t-test.Results The expression level of SP1 protein in adjacent tissues of cancer(1.03±0.22)was significantly lower than that in ovarian cancer tissue(1.93±0.39,t=17.570,P<0.05).The absorbance(A)value of the control group(1.98±0.09)was significantly higher than that of the SP KD1 group(1.27±0.14,t=10.310,P<0.05).The EdU positive rate of the control group[(84.17±4.07)%]was significantly higher than that of the SP KD1 group[(65.67±5.57)%,t=6.566,P<0.05].The scratch healing rate of the control group[(86.46±3.94)%]was significantly higher than that of the SP KD1 group[(74.25±5.07)%,t=4.659,P<0.05].The number of invasive cells in the control group[(139.67±12.09)cells]was significantly greater than that in the SP KD1 group[(97.67±9.52)cells,t=6.684,P<0.05].The interstitial cell markers N-cadherin and Vimentin(0.88±0.08;1.12±0.09)in the control group were significantly higher than those in the SP KD1 group(0.68±0.08;0.78±0.07,t=4.512,7.335,P<0.05).The epithelial cell marker E-cadherin(0.81±0.06)in the control group was significantly lower than that in the SP KD1 group(1.25±0.14,t=7.156,P<0.05).The expression level of microRNA(miR)-182 in the control group(1.10±0.15)was significantly higher than that in the SP KD1 group(0.79±0.08,t=4.503,P<0.05).Conclusion Transcription factor SP1 is highly expressed in ovarian cancer tissue,which promotes the proliferation and migration ability of ovarian cancer cells by increasing the expression level of miR-182.