摘要
目的探究甾醇调节元件结合蛋白1(SREBP1)在宫颈上皮内瘤变(CIN)与宫颈癌(CC)中的表达变化及其对肿瘤进程的影响。方法收集CIN组织与CC组织各20例,免疫组化染色检测SREBP1阳性表达,实时荧光定量PCR反应(RT-qPCR)测定SREBP1mRNA表达水平;将人宫颈癌HeLa细胞分为对照组、pcDNA3.1-NC组、pcDNA3.1-SREBP1组、sh-NC组、sh-SREBP1组,CCK-8法检测各组细胞增殖活性,细胞克隆形成实验检测各组细胞克隆形成数目,免疫荧光染色观察各组细胞内上皮型钙黏蛋白(E-cadherin)表达,体外小管形成实验检测各组细胞成管情况,Western blot测定各组细胞内E-cadherin、神经型钙黏蛋白(N-cadherin)、波形蛋白(Vimentin)、基质金属蛋白酶-9(MMP-9)和血管内皮生长因子(VEGF)蛋白表达。结果随着CIN分级程度的增加,SREBP1表达越高,且CC组织SREBP1表达显著高于CIN组织(P<0.05)。与pcDNA3.1-NC组比较,pcDNA3.1-SREBP1组细胞增殖活性升高,克隆形成数目增加,细胞内E-cadherin荧光表达明显减弱,血管管腔形成数目增多,E-cadherin蛋白表达下调,N-cadherin、Vimentin、MMP-9及VEGF蛋白表达均上调(P<0.05)。与sh-NC组比较,sh-SREBP1组细胞增殖活性降低,克隆形成数目减少,细胞内E-cadherin荧光表达则明显增强,同时,血管管腔形成数目减少,E-cadherin蛋白表达上调而N-cadherin、Vimentin、MMP-9、VEGF蛋白表达均下调(P<0.05)。结论SREBP1在CIN与CC组织中表达升高,抑制其表达能够降低CC肿瘤细胞增殖,阻止肿瘤细胞上皮间质转化与血管形成,从而减缓肿瘤进程。
Objective To investigate the expression changes of sterol regulatory element-binding protein 1(SREBP1)in cervical intraepithelial neoplasia(CIN)and cervical cancer(CC)and its effect on tumor progression.Methods Twenty cases of CIN tissues and CC tissues were collected,and the positive expression of SREBP1 was detected by immunohistochemical staining,the expression level of SREBP1 mRNA was determined by real-time fluorescence quantitative PCR reaction(RT-qPCR).Human cervical cancer HeLa cells were divided into control group,pcDNA3.1-NC group,pcDNA3.1-SREBP1 group,sh-NC group and sh-SREBP1 group,CCK-8 method was used to detect the proliferation activity of cells in each group,the cell clone formation experiment was used to detect the number of cell clones formed in each group,immunofluorescence staining was used to observe the expression of epithelial cadherin(E-cadherin)in the cells of each group,in vitro tubule formation experiments were used to detect the tube formation of cells in each group.Western blot was used to detect the expressions protein of E-cadherin,N-cadherin,Vimentin,matrix metalloproteinase-9(MMP-9)and vascular endothelial growth factor(VEGF)in each group.Results With the increase of CIN grade,the expression of SREBP1 was higher,and the expression of SREBP1 in CC tissue was significantly higher than that in CIN tissue(P<0.05).Compared with the pcDNA3.1-NC group,the cell proliferation activity of the pcDNA3.1-SREBP1 group increased,the number of cell clones increased,the intracellular E-cadherin fluorescence expression was significantly reduced,the number of vascular lumen formation increased,the protein expressions of E-cadherin were down-regulated,the protein expressions of N-cadherin,Vimentin,MMP-9 and VEGF were all up-regulated(P<0.05).Compared with the sh-NC group,the cell proliferation activity of the sh-SREBP1 group was reduced,the number of cell clones was reduced,the intracellular E-cadherin fluorescence expression was significantly enhanced,at the same time,the number of vascular lumen formed was decreased,the expression of E-cadherin protein was up-regulated,while the protein expressions of N-cadherin,Vimentin,MMP-9 and VEGF were down-regulated(P<0.05).Conclusion The expression of SREBP1 is increased in CIN and CC tissues,and inhibiting its expression can reduce the proliferation of CC tumor cells,prevent the epithelial-mesenchymal transition and angiogenesis of tumor cells,thereby slowing the tumor progression.
作者
阿依努尔·买苏提
迪丽努尔·安外尔
陆萍
Ayinur Maisuti;Dilinur Anwaier;LU Ping(Department of Gynecology,People’s Hospital of Xinjiang Uygur Autonomous Region,Urumqi,Xinjiang 830001,China)
出处
《中国优生与遗传杂志》
2023年第6期1102-1108,共7页
Chinese Journal of Birth Health & Heredity
基金
新疆维吾尔自治区自然科学基金(2021D01C219)。
关键词
宫颈上皮内瘤变
宫颈癌
甾醇调节元件结合蛋白1
上皮间质转化
血管形成
cervical intraepithelial neoplasia
cervical cancer
sterol regulatory element-binding protein 1
epithelial-mesenchymal transition
angiogenesis
