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目的:检测酰基-辅酶A去饱和酶1(stearoyl-CoA desaturase-1,SCD-1)在食管鳞癌组织和细胞中的表达,分析其表达下调对食管鳞癌EC1细胞增殖和凋亡的影响,并探讨其相关的分子机制。方法:采用免疫组织化学和原位杂交法检测60例食管鳞癌组织及其相应癌旁正常食管黏膜组织中SCD-1mRNA和蛋白的表达,实时荧光定量PCR和蛋白质印迹法检测正常食管组织(作为阴性对照)和食管鳞癌细胞株中SCD-1mRNA和蛋白的表达。不同浓度SCD-1小干扰RNA(small interfering RNA,siRNA)转染食管鳞癌EC1细胞后,采用实时荧光定量PCR和蛋白质印迹法检测SCD-1mRNA和蛋白的表达,应用CCK-8法检测转染前后EC1细胞增殖的变化,FCM检测转染前后EC1细胞凋亡的改变,蛋白质印迹法检测总Akt、p-Akt、bcl-2和bax蛋白的表达。结果:食管鳞癌组织中SCD-1mRNA和蛋白表达的阳性率显著高于正常食管黏膜组织(P<0.05),其表达上调与肿瘤组织分级、TNM分期和淋巴结转移密切相关(P<0.05)。此外,与正常食管组织相比,食管鳞癌EC9706、Eca109和EC1细胞中SCD-1mRNA和蛋白表达均显著上调(P<0.05),其中EC1细胞中SCD-1的表达水平最高。50nmol/LSCD-1siRNA能显著下调EC1细胞中SCD-1mRNA和蛋白的表达。SCD-1表达下调可显著抑制EC1细胞的增殖,诱导细胞凋亡;同时,显著上调bax蛋白的表达,并下调p-Akt和bcl-2蛋白的表达,但不改变总Akt的表达水平。结论:SCD-1可能在食管鳞癌的发生和发展中具有重要作用,抑制SCD-1的表达有望成为食管鳞癌重要的分子治疗策略之一。
OBJECTIVE: To detect the expression of stearoyl-CoA desaturase-1 (SCD-1) in esophageal squamous cell carcinoma tissues and cells, and to investigate the effect of its expression down-regulation on the proliferation and apoptosis of esophageal squamous cell carcinoma EC1 cells , And explore its related molecular mechanisms. Methods: Immunohistochemistry and in situ hybridization were used to detect the expression of SCD-1 mRNA and protein in 60 cases of esophageal squamous cell carcinoma and its adjacent normal esophageal mucosa. Real-time fluorescence quantitative PCR and Western blotting were used to detect the expression of SCD- Negative control) and esophageal squamous cell carcinoma SCD-1 mRNA and protein expression. The expression of SCD-1 mRNA and protein was detected by real-time fluorescence quantitative PCR and Western blotting after transfection of small interfering RNA (siRNA) with different concentrations of SCD-1 into EC1 cells. CCK-8 was used to detect the expression of SCD- The changes of EC1 cell proliferation were detected by FCM. The expressions of Akt, p-Akt, bcl-2 and bax proteins were detected by Western blotting. Results: The positive rate of SCD-1 mRNA and protein expression in esophageal squamous cell carcinoma was significantly higher than that in normal esophageal mucosa (P <0.05). The up-regulation of SCD-1 mRNA and protein expression was closely related to tumor grade, TNM stage and lymph node metastasis (P <0.05). In addition, SCD-1 mRNA and protein expressions in esophageal squamous cell carcinoma EC9706, Eca109 and EC1 cells were significantly up-regulated compared with normal esophageal tissues (P <0.05), of which SCD-1 expression was the highest in EC1 cells. 50 nmol / LSCD-1 siRNA significantly down-regulated the expression of SCD-1 mRNA and protein in EC1 cells. The down-regulation of SCD-1 can significantly inhibit the proliferation and induce the apoptosis of EC1 cells. At the same time, down-regulation of bax protein expression and down-regulation of p-Akt and bcl-2 protein expression did not change the expression of total Akt. Conclusion: SCD-1 may play an important role in the development and progression of esophageal squamous cell carcinoma. Suppression of SCD-1 expression may be one of the important molecular therapy strategies for esophageal squamous cell carcinoma.