下调胰岛素样生长因子-1表达对人胃癌细胞增殖和侵袭及迁移的影响

发布时间：2018-04-04 03:25

本文选题：胰岛素样生长因子-　切入点：shRNA　出处：《中华肿瘤防治杂志》2017年01期

【摘要】：目的胰岛素样生长因子-1(insulin like growth factor-1,IGF-1)作为体内重要的促有丝分裂原分子,可能与恶性肿瘤的转化及转移相关,但是目前有关IGF-1的表达与胃癌细胞迁移、侵袭的研究较少。本研究旨在研究IGF-1在胃癌组织中的表达,并探讨其表达对人胃癌HGC-27细胞体外增殖及侵袭、迁移能力的影响。方法检测承德医学院附属医院2014-05-10-2016-05-10胃癌手术切除78例胃癌组织、30例癌旁正常组织及体外不同胃癌细胞系中IGF-1的表达情况。针对IGF-1基因序列,设计发夹RNA(short-hairpin RNA,shRNA)干扰序列并构建干扰载体,转染胃癌HGC-27细胞。荧光实时定量PCR及蛋白质印迹技术分别检测转染干扰载体后HGC-27细胞中IGF-1的表达情况。细胞增殖-毒性检测试剂盒(cell counting kit-8,CCK-8)、划痕实验及Transwell侵袭实验分别检测下调IGF-1对体外细胞HGC-27增殖及迁移、侵袭能力的影响。结果免疫组织化学法检测显示,低分化胃癌组织中IGF-1阳性表达率为76%(29/38),高、中分化为53%(21/40),癌旁正常组织为27%(8/30)。不同胃癌组织与癌旁组织中IGF-1的表达差异有统计学意义,χ2=16.66,P0.01。并且其表达与胃癌的病理分期(χ2=7.430,P=0.007)、组织分化(χ2=4.618,P=0.032)等病理因素相关。体外不同胃癌细胞系中IGF-1均呈现高表达,但差异无统计学意义。利用干扰载体能够有效下调胃癌细胞HGC-27中IGF-1在mRNA及蛋白质水平的表达。与空载体转染对照组和野生型HGC-27细胞比较,IGF-1表达下调明显抑制胃癌细胞的体外增殖和侵袭、迁移能力。结论 IGF-1在体内胃癌组织及体外胃癌细胞中高表达,下调IGF-1的表达能够抑制人胃癌细胞HGC-27的增殖及体外侵袭、迁移能力。
[Abstract]:Objective Insulin-like growth factor-1insulin like growth factor-1 (IGF-1), as an important mitogen molecule in vivo, may be related to the transformation and metastasis of malignant tumors, but there are few studies on the expression of IGF-1 and the migration and invasion of gastric cancer cells.The purpose of this study was to investigate the expression of IGF-1 in gastric carcinoma and its effect on the proliferation, invasion and migration of human gastric cancer HGC-27 cells in vitro.Methods the expression of IGF-1 in gastric cancer tissues from 78 cases of gastric cancer tissues and 30 cases of adjacent normal tissues and different gastric cancer cell lines in vitro were detected in 2014-05-10-2016-05-10 gastric cancer surgery in affiliated Hospital of Chengde Medical College.Aimed at the IGF-1 gene sequence, the interference sequence of hairpin RNA(short-hairpin RNA-shRNAs was designed and the interference vector was constructed to transfect gastric cancer HGC-27 cells.The expression of IGF-1 in HGC-27 cells after transfection of interference vector was detected by real-time quantitative PCR and Western blot respectively.Cell counting kit-8 CCK-8, scratch test and Transwell invasion assay were used to detect the effect of down-regulation of IGF-1 on proliferation, migration and invasion of HGC-27 in vitro.Results Immunohistochemical assay showed that the positive expression rate of IGF-1 in poorly differentiated gastric cancer tissues was 7629 / 38%, which was high and differentiated into 5321 / 40%, and 27 / 8 / 30% in adjacent normal tissues.The expression of IGF-1 in different gastric cancer tissues and paracancerous tissues was significantly different (蠂 ~ 2 ~ (2) ~ (16) 6 ~ (6)) P ~ (0.01).And its expression was correlated with pathological stage of gastric cancer (蠂 ~ 2 ~ 2 ~ (7.430) P ~ (0.007)) and histological differentiation (蠂 ~ (2) ~ (2) ~ (4.618) P ~ (0.032)).High expression of IGF-1 was found in different gastric cancer cell lines in vitro, but the difference was not statistically significant.The expression of IGF-1 in mRNA and protein in gastric cancer cell line HGC-27 can be effectively down-regulated by interference vector.Compared with the control group and wild-type HGC-27 cells transfected with empty vector, the down-regulated expression of IGF-1 significantly inhibited the proliferation, invasion and migration of gastric cancer cells in vitro.Conclusion IGF-1 is highly expressed in gastric cancer tissues in vivo and in vitro, and down-regulation of IGF-1 expression can inhibit the proliferation, invasion and migration of human gastric cancer cells HGC-27 in vitro.
【作者单位】：承德医学院附属医院临床技能教学中心;承德医学院附属医院病理科;承德医学院附属医院老年病科;承德医学院附属医院手足外科;
【分类号】：R735.2

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