胃腺癌细胞中血管活性肠肽自分泌调节作用及其机制  

The autocrine route of vasoactive intestinal peptide in gastric adenocarcinoma cell line and its mechanism

作　　者：李国华[1] 钱伟[1] 王静[1] 侯晓华[1] 

机构地区：[1]华中科技大学同济医学院附属协和医院消化科

出　　处：《中华消化杂志》2005年第1期27-31,共5页Chinese Journal of Digestion

摘　　要：目的　研究胃腺癌细胞株SGC7901细胞中是否存在血管活性肠肽(VIP)的自分泌调节作用及其机制。方法　用RT PCR检测SGC7901细胞中VIP及其受体(VIPR1、VIPR2 )mRNA的表达,用免疫组化观察SGC7901细胞蛋白的表达,用放射免疫法测定细胞上清液中VIP含量,用MTT及细胞计数法观察外源VIP和VIP受体拮抗剂 [D p Cl Phe6, Leu17 ] VIP对SGC7901细胞增殖的影响。同时用RT PCR检测外源VIP及其受体拮抗剂孵育前后细胞c myc、鸟氨酸脱羧酶(ODC)mRNA的表达量变化。结果　SGC7901细胞不仅表达VIPmRNA,而且是VIP免疫反应阳性细胞,VIP免疫反应阳性颗粒主要分布在细胞质,少数在细胞核。细胞培养上清液中可检测到VIP蛋白,平均每 106 个细胞可分泌 (13. 15±8. 54)pg的VIP。SGC7901细胞也能表达VIPR1 和VIPR2 mRNA。10-5 ~10-12 mol/L外源性VIP孵育 24h后,细胞增殖无显著影响 (P>0. 05),而 10-5 ~10-8 mol/L拮抗剂显著促进SGC7901细胞增殖(P<0. 05)。10-6 mol/L外源性VIP孵育 24h后,SGC7901细胞c myc及ODCmRNA的表达量显著下降(RM /β: 0. 816±0. 049比 0. 901±0. 054,P<0. 01;RO/β: 0. 737±0. 060比 0. 830±0. 051,P<0. 01);而10-6 mol/L[D p Cl Phe6, Leu17 ] VIP使细胞c myc及ODCmRNA的表达量显著升高 (RM/β: 0.Objective To investigate whether there is autocrine route of vasoactive intestinal peptide (VIP) in SGC7901 gastric adenocarcinoma cell line, and how the autocrine may affect the growth of SGC7901 cell. Methods VIP and its receptors (VIPR_1, VIPR_2) mRNA in SGC7901 cells were detected by RT-PCR, VIP protein in cells by immunohistochemistry, and VIP protein in supernatant of cells by radioimmunoassay. The effect of exogenous VIP and its antagonist [D-p-Cl-Phe6, Leu17]-VIP on the amplification of SGC7901 cells were observed by MTT and cell count. Expression quantity alteration of c-myc mRNA and ornithine decarboxylase (ODC) mRNA by RT-PCR after incubation with exogenous VIP or [D-p-Cl-Phe6, Leu17]-VIP were detected. Results SGC7901 cells not only expressed VIP mRNA, but also were positive reactive to VIP. Most of the positive particls distributed in cytoplasm, but few in karyon. 10~6 SGC7901 cells could secrete (13.15(±8.54)pg) VIP protein on average in supernatant. SGC7901 cells could also express VIP receptors mRNA. Exogenous VIP could not affect the growth of SGC7901 cells incubated in 10^(-5)-10^(-12) mol/L for 24 h(P>0.05). However, 10^(-5)-10^(-8) mol/L antagonist facilitated the growth of the cells(P<0.05). Exogenous VIP incubated in 10^(-6) mol/L for 24 h inhibited the expressions of c-myc and ODC mRNA (R_(M/β):(0.816)±0.049 vs (0.901)±0.054; and R_(O/β):0.737±0.060 vs 0.830±0.051 respectively, P<0.01), but 10^(-6) mol/L [D-p-Cl-Phe6, Leu17]-VIP incubated for 24 h enhanced their expressions (R_(M/β):0.950±(0.045) vs 0.901±0.054, P<(0.05;) and R_(O/β): 0.911±0.062 vs 0.830±0.051, P<0.01 respectively). Conclusions There is autocrine route of VIP in SGC7901 cell line, which inhibits their growth by inhibiting c-myc and ODC mRNA expression.

关 键 词：VIP SGC7901细胞 RNA 表达量 自分泌 血管活性肠肽 调节作用 胃腺癌细胞 细胞核 蛋白 

分 类 号：R735.2[医药卫生—肿瘤]