TRPV6对结肠癌SW480细胞生物学行为及小鼠模型中结肠肿瘤发生的影响  

Effect of TRPV6 channels on the biological behavior of sw480 colon cancer cell line and colonic tumorigenesis in rat model

作　　者：谢臣武 刘模荣[1] 董辉[2] 徐靖宇[1] 谢睿[1] 文国荣[1] 刘莲花[1] Chenwu Xie;Morong Liu;Hui Dong;Jingyu Xu;Rui Xie;Guorong Wen;Lianhua Liu(Department of Gastroenterology,Affiliated Hospital,Zunyi Medical College,Zunyi 563003,China;Division of Gastroenterology,Department of Medicine,University of California,San Diego,School of Medicine,San Diego 92103,USA)

机构地区：[1]遵义医学院附属医院消化内科,贵州省遵义市563003 [2]美国加利福尼亚大学医学院胃肠科

出　　处：《中国肿瘤临床》2018年第21期1079-1085,共7页Chinese Journal of Clinical Oncology

基　　金：贵州省国际科技合作计划项目[编号:黔科合G字(2014)7014]资助~~

摘　　要：目的:通过观察沉默瞬时受体阳离子通道亚家族V成员6(transient receptor potential cation channel,subfamily V member 6,TRPV6)对结肠癌SW480细胞生物学行为的影响和细胞内钙的浓度变化,以及1,25(OH)_2D_3、CaCl_2及CuCl_2在SD大鼠结肠肿瘤模型建立中的影响,探索TRPV6在结肠癌发生过程中的相关作用,为结肠癌的防治寻找新的靶点。方法:通过构建慢病毒颗粒感染结肠癌SW480细胞,采用免疫组织化学法、Western bolt、PCR技术检测TRPV6蛋白及m RNA的表达,MTT法、迁移及凋亡实验观察结肠癌SW480细胞增殖、迁移及凋亡变化,高速离子成像系统测定结肠癌SW480细胞内的Ca^(2+)浓度变化。以二甲基肼(dimethyl hydrazine,DMH)建立SD大鼠结肠肿瘤模型,分为实验组(DMH组)和干预组[(DMH+1,25(OH)_2D_3组、DMH+CuCl_2组)]、对照组,干预组分别予1,25(OH)_2D_3(37.5 nmol/kg)、CuCl2(375μmol/kg)对模型进行干预。观察各组大鼠结肠腺瘤及腺癌的发生情况,Western blot检测各组结肠组织中TRPV6蛋白的表达情况。结果:TRPV6-RNAi转染结肠癌SW480细胞后,TRPV6 m RNA及蛋白表达减少,细胞Ca^(2+)浓度水平降低,结肠癌SW480细胞的增殖率、迁移能力下降,细胞凋亡率增加,与空白对照组及阴性对照组比较差异具有统计学意义(P<0.05)。对照组大鼠成瘤率为0,DMH+1,25(OH)_2D_3组为100%,DMH组为84.62%,DMH+CuCl_2组为33.33%。各组大鼠大肠中均有TRPV6蛋白的表达,表达情况为DMH+1,25(OH)_2D_3组>DMH组>DMH+CuCl_2组>对照组,差异具有统计学意义(P<0.05)。结论:降低结肠癌SW480细胞Ca^(2+)浓度水平可抑制结肠癌SW480细胞的增殖、迁移能力,诱导细胞凋亡。1,25(OH)_2D_3可以增加实验鼠结肠组织TRPV6蛋白表达,促进结肠肿瘤的形成。CuCl_2可以使实验鼠结肠组织中的TRPV6蛋白表达降低,并抑制结肠肿瘤的形成。Objective： To observe the influence of TRPV6 gene silence on SW480 colon cancer cell biological behavior, change in the intracellular concentration of calcium, as well as influence of 1,25（OH）2D3 CaCl2 and CuCl2 on SD rat colonic neoplasm models. Method： SW480 colon cancer cells were infected using lentivirus particles. TRPV6 protein and m RNA expression was detected using immunohistochemical tests, Western blot, and PCR. Moreover, the proliferation, metastasis, and apoptosis of SW480 colon cancer cells were detected through MTT assay and metastasis and apoptosis experiments, and the concentration of Ca2＋ in SW480 colon cancer cells was measured using highspeed ionic imaging. The SD rat colon cancer model was established based on DMH, and were assigned into experimental group（DMH group,15） and intervention group（DMH＋1,25（OH）2D3 group, DMH＋CuCl2 group） and control group, 10 in each group. The SD rat colon cancer model is established based on DMH, given 1,25（OH）2D3（37.5 nmol/kg） and CuCl2（375 μmol/kg） separately as intervention. The occurrence of colonic neoplasms and glandular cancers in each group of rats was observed, and Western blot was employed for detection of the TRPV6 protein expression. Results： After the transfection of SW480 colon cancer cells by TRPV6-RNAi, the expression of TRPV6 m RNA and protein decreased,intracellular concentration of Ca2＋decreased, proliferation and metastasis rate of SW480 colon cancer cells decreased, and apoptosis rate of these cells increased. The differences between the groups with intervention and the blank control group and negative control group showed statistical significance（P〈0.05）. The colon cancer occurrence rate in the control group was 0, while that of the DMH＋1,25（OH）2 D3 group, DMH group, and DMH＋CuCl2 were 100%, 84.62%, and 33.33%, respectively. The TRPV6 protein expression was detected in all groups,while DMH＋1,25（OH）2D3 group was observed to exhibit the highest level of expression, followed by the D

关 键 词：TRPV6 基因沉默 钙离子通道 结肠癌 二甲基肼 

分 类 号：R735.35[医药卫生—肿瘤]