FDFT1依赖肿瘤免疫微环境促进结肠癌细胞的生长  

Farnesyl-diphosphate farnesyltransferase 1 promotes growth of colorectal cancer cells with dependence on tumor immune microenvironment

作　　者：罗娜 张大鹏[2] 缪洪明 代荣阳[1] LUO Na;ZHANG Dapeng;MIAO Hongming;DAI Rongyang(College of Basic Medical Sciences,Southwest Medical University,Luzhou,Sichuan Province,646000;Department of Biochemistry and Molecular Biology,College of Basic Medical Sciences,Army Medical University(Third Military Medical University),Chongqing,400038,China)

机构地区：[1]西南医科大学基础医学院,四川泸州646000 [2]陆军军医大学(第三军医大学)基础医学院生物化学与分子生物学教研室,重庆400038

出　　处：《第三军医大学学报》2021年第24期2648-2655,共8页Journal of Third Military Medical University

基　　金：国家自然科学基金面上项目(81872028)。

摘　　要：目的研究法呢基二磷酸酯法呢基转移酶1 (farnesyl-diphosphate farnesyltransferase 1,FDFT1)对结肠癌(colorectal cancer,CRC)细胞MC-38体内体外生长的影响。方法通过GEPIA数据库分析275例CRC样本和349例正常组织样本中FDFT1的表达量,通过数据库分析人CRC中FDFT1的免疫组化表达水平。使用慢病毒介导的短发夹RNA(short hairpin RNA,shRNA)构建FDFT1沉默的CRC细胞系MC-38,得到稳定低表达FDFT1的MC-38细胞系(sh-FDFT1组)和空载对照细胞系(sh-NC组),Western blot、RT-qPCR验证敲低效果。CCK-8实验、流式细胞术、Transwell实验分别观察敲低FDFT1对MC-38细胞增殖、凋亡、迁移能力的影响。采用C57BL/6小鼠、重度免疫缺陷NCG小鼠构建CRC腹腔种植转移模型,利用C57BL/6小鼠构建皮下移植瘤模型,14 d后观察沉默FDFT1对小鼠肿瘤生长情况的影响。结果数据库分析结果显示CRC组织中FDFT1的表达较正常组织高(P<0.05),免疫组化结果显示CRC病人FDFT1高表达。沉默FDFT1对MC-38细胞增殖、凋亡及迁移均没有明显影响;在C57BL/6小鼠CRC腹腔种植转移模型和皮下移植瘤模型中均发现沉默FDFT1后显著抑制肿瘤生长(P<0.05)。NCG小鼠腹腔种植转移模型显示沉默FDFT1后肿瘤生长没有差异(P>0.05)。结论 FDFT1在体内促进CRC细胞生长依赖于肿瘤免疫微环境。Objective To investigate the effects of farnesyl-diphosphate farnesyltransferase 1(FDFT1) on the growth of colorectal cancer(CRC) cell line MC-38 in vitro and in mouse models of colorectal peritoneal metastasis and xenograft tumor.Methods The expression of FDFT1 in 275 CRC samples and 349 normal tissue samples,as well as the immunohistochemical expression level of FDFT1 in human CRC tissues were analyzed using online databases Gene Expression Profiling Interactive Analysis(GEPIA) and the Human Protein Atlas,respectively.FDFT1-silenced CRC cell line MC-38 was constructed by lentivirus-mediated short hairpin RNA(shRNA) to obtain stable cells with low expression of FDFT1(sh-FDFT1),and blank control cells(sh-NC) was also constructed.The knockdown effect was subsequently verified by Western blotting and RT-qPCR.Cell counting kit-8(CCK-8) assay,flow cytometry and transwell assay were performed to observe the effects of FDFT1 knockdown on the proliferation,apoptosis and migration of MC-38 cells,respectively.In addition,intraperitoneal implantation models of colorectal cancer were established using C57 BL/6 mice and severe immunodeficient NCG mice,and subcutaneous transplantation tumor model was also constructed using C57 BL/6 mice.The effects of FDFT1 silencing on tumor growth in vivo were observed in 14 d after transplantation.Results Database analysis showed that FDFT1 expression was higher in CRC tissues than normal tissues(P<0.05),and immunohistochemical results also confirmed that FDFT1 was highly expressed in CRC tissues.There were no significant alterations in the proliferation,apoptosis and migration of MC-38 cells after knockdown of FDFT1.Although FDFT1 silencing obviously inhibited the tumor growth in both intraperitoneal implantation and subcutaneous xenograft models of CRC in C57 BL/6 mice(P<0.05),the colorectal peritoneal metastasis model of NCG mice presented no difference in tumor growth after silencing FDFT1(P>0.05).Conclusion FDFT1 depends on the tumor immune microenvironment in the promotion of the gro

关 键 词：法呢基二磷酸酯法呢基转移酶1 结肠癌 免疫微环境 MC-38 

分 类 号：R345[医药卫生—基础医学] R730.23