摘要
[目的]研究FAT4通过上调乳腺癌耐药蛋白(BCRP)影响结直肠癌对5-FU的耐药性。[方法]采用RT-PCR检测50例结直肠癌患者中癌组织以及相应癌旁组织和正常组织中FAT4表达量;采用免疫组化检测手术组以及手术+新辅助化疗组中FAT4以及BCRP的相对表达量,并采用Pearson相关性分析FAT4与BCRP表达水平间的相关性;采用RT-PCR检测各结直肠癌细胞系(SW48、HT-29、HCT-116、SW620、DLD1、SW480、CaCO2、LOVO)中FAT4和BCRP mRNA的表达量;构建过表达FAT4或敲降FAT4的直肠癌细胞系,用RT-PCR以及Western blot检测FAT4表达水平的变化;进一步使用5-FU刺激过表达FAT4或敲降FAT4的直肠癌细胞系,MTT检测其增殖能力的变化。[结果]与正常组织以及癌旁组织比较,FAT4以及BCRP在癌组织中表达量较高(P<0.001),5-FU刺激结直肠癌细胞后,与对照组比较,FAT4过表达可以增加其细胞增殖能力(P<0.01),FAT4敲减可以降低细胞增殖能力(P<0.01)。Pearson相关性分析显示,结直肠癌患者中FAT4和BCRP表达水平存在明显的正相关(P<0.001)。在HT-29细胞中FAT4过表达能促进BRCP的表达,在SW480细胞中FAT4敲减后会抑制BRCP的表达;在过表达FAT4的结直肠癌细胞中转染shBCRP会降低其对5-FU的耐药性。[结论]FAT4通过上调耐药基因BCRP,使直肠癌细胞对5-FU的耐药性增加。
[Objective]To study the effect of FAT4 on 5-FU resistance of colorectal cancer by upregulation of breast cancer resistance protein(BCRP).[Methods]The expression of FAT4 in 50 patients with colorectal cancer was detected by RT-PCR;The relative expressions of FAT4 and BCRP in the operation group and the operation+neoadjuvant chemotherapy group were detected by immunohistochemistry,and the correlation between FAT4 and BCRP expression levels was analyzed by Pearson correlation analysis.The expressions of FAT4 and BCRP mRNA in colorectal cancer cell lines(SW48,ht-29,hct-116,SW620,DLD1,SW480,CaCO2,LOVO)were detected by RT-PCR.Colorectal cancer cell lines overexpressing FAT4 or knockout of FAT4 were constructed,and the changes of FAT4 expression levels were detected by RT-PCR and Western blot.5-FU was further used to stimulate colorectal cancer cell lines that overexpressed FAT4 or knockout of FAT4,and MTT was used to detect changes in their proliferation capacity.[Results]Compared with normal tissues and adjacent tissues,FAT4 and BCRP were highly expressed in cancer tissues(P<0.001).After 5-FU stimulation of colorectal cancer cells,FAT4 overexpression could increase the proliferation ability of cells(P<0.01),while FAT4 knockdown could decrease the proliferation ability of cells(P<0.01).Pearson correlation analysis showed that there was a significant positive correlation between the expression levels of FAT4 and BCRP in colorectal cancer patients(P<0.001).Overexpression of FAT4 in HT-29 cells promoted the expression of BRCP,while knockdown of FAT4 in SW480 cells inhibited the expression of BRCP.Co-transfection of shBCRP in colorectal cancer cells overexpressing FAT4 reduced their resistance to 5-FU.[Conclusion]FAT4 increased resistance to 5-FU in colorectal cancer cells by upregulating BCRP.
作者
张心武
张寅斌
张迪
李顺乐
孙晓力
雷焕芹
翟宏军
李华
ZHANG Xin-wu;ZHANG Yin-bin;ZHANG Di;LI Shun-le;SUN Xiao-li;LEI Huan-qin;ZHAI Hong-jun;LI Hua(Department of General Surgery,the Second Affiliated Hospital of Xi'an Jiao Tong University,Xi'an 710004,China;Department of Oncology,the Second Affiliated Hospital of Xi'an Jiao Tong University,Xi'an 710004,China)
出处
《中国中西医结合消化杂志》
CAS
2020年第7期508-513,共6页
Chinese Journal of Integrated Traditional and Western Medicine on Digestion
基金
西安交通大学基本科研业务(No:Xjj2012072)。
