Zhang D J, Xiang J, Wang X, Wang J, Xiao J C, Xu W, Xu H, Xin Y, Zhang L Z, Pei D S, Zheng J N, Gu Y M
Department of Interventional Radiology Affiliated Hospital of Xuzhou Medical College Xuzhou, Jiangsu, China -
Panminerva Med. 2015 Dec;57(4):183-9.
To determinate the RPA1 expression in esophageal carcinoma and the paired tumor-adjacent tissue, and to explore the influence of RPA1 on radiosensitivity of esophageal carcinoma TE-1 cells.
Firstly, the RPA1 expression of 40 cases esophageal carcinoma and their adjacent tissues were detected by immunohistochemistry. Secondly, The esophageal carcinoma cell subline-radiation resistance model (TE-1R) was constructed by radiation-induction, the RPA1 expression and proliferation activity of TE-1 and TE-1R cells were detected by Western blot and MTT assay respectively. After radiation, the expression of RPA1 and cell apoptosis were detected by Western blot and FACS respectively. Cell clone formation and survival rate were detected by clonogenic assay. Thirdly, Inhibiting RPA1 expression by siRNA in TE-1 cells, the expression of RPA1 was detected by RT-PCR and Western blot, Cell proliferation inhibition ratio and cell apoptosis after radiation were detected by MTT assay and FACS respectively.
The RPA1 expression in esophageal carcinoma was significantly higher than that in the tumor-adjacent tissues, which was associated with tumor invasion and lymph node metastasis. The RPA1 expression in TE-1R cells was higher than that in TE-1 cells, while the proliferation activity of TE-1R cells was lower than that of TE-1 cells, and the apoptosis rate of TE-1R cells after radiation was less than that of TE-1 cells. In addtion, the clone formation and survival rate of TE-1R cells were higher than that of TE-1 cells. Moreover, inhibiting RPA1 expression by siRNA-RPA1 could promoted proliferation inhibition ratio and apoptosis rate of TE-1 cells after radiation.
The over-expression of RPA1 in esophageal carcinoma was related with progression and metastasis. Moreover, radiation induced the excessive expression RPA1 in TE-1 cells, and the radiosensitivity of TE-1R cells was less than that of TE-1 cells. Furthermore, inhibiting RPA1 expression could increase radiosensitivity of TE-1 cells. Overall, RPA1 could influence radiosensitivity and might be one important mechanism of radiation resistance in TE-1 cells.
检测食管癌及配对癌旁组织中RPA1的表达,探讨RPA1对食管癌TE-1细胞放射敏感性的影响。
首先,采用免疫组织化学法检测40例食管癌及其癌旁组织中RPA1的表达。其次,通过辐射诱导构建食管癌细胞亚系辐射抗性模型(TE-1R),分别采用蛋白质免疫印迹法和MTT法检测TE-1和TE-1R细胞中RPA1的表达及增殖活性。辐射后,分别采用蛋白质免疫印迹法和流式细胞术检测RPA1的表达及细胞凋亡情况。采用克隆形成实验检测细胞克隆形成及存活率。第三,在TE-1细胞中通过小干扰RNA(siRNA)抑制RPA1表达,采用逆转录聚合酶链反应(RT-PCR)和蛋白质免疫印迹法检测RPA1的表达,分别采用MTT法和流式细胞术检测辐射后细胞增殖抑制率及细胞凋亡情况。
食管癌中RPA1的表达明显高于癌旁组织,且与肿瘤侵袭及淋巴结转移相关。TE-1R细胞中RPA1的表达高于TE-1细胞,而TE-1R细胞的增殖活性低于TE-1细胞,且辐射后TE-1R细胞的凋亡率低于TE-1细胞。此外,TE-1R细胞的克隆形成及存活率高于TE-1细胞。而且,通过siRNA-RPA1抑制RPA1表达可提高辐射后TE-1细胞的增殖抑制率及凋亡率。
食管癌中RPA1的过表达与肿瘤进展及转移相关。此外,辐射诱导TE-1细胞中RPA1过度表达,且TE-1R细胞的放射敏感性低于TE-1细胞。而且,抑制RPA1表达可提高TE-1细胞的放射敏感性。总体而言,RPA1可影响放射敏感性,可能是TE-1细胞辐射抗性的重要机制之一。