本文選題：胰島素樣生長因子-　切入點：shRNA　出處：《中華腫瘤防治雜志》2017年01期

【摘要】：目的胰島素樣生長因子-1(insulin like growth factor-1,IGF-1)作為體內(nèi)重要的促有絲分裂原分子,可能與惡性腫瘤的轉(zhuǎn)化及轉(zhuǎn)移相關(guān),但是目前有關(guān)IGF-1的表達與胃癌細胞遷移、侵襲的研究較少。本研究旨在研究IGF-1在胃癌組織中的表達,并探討其表達對人胃癌HGC-27細胞體外增殖及侵襲、遷移能力的影響。方法檢測承德醫(yī)學(xué)院附屬醫(yī)院2014-05-10-2016-05-10胃癌手術(shù)切除78例胃癌組織、30例癌旁正常組織及體外不同胃癌細胞系中IGF-1的表達情況。針對IGF-1基因序列,設(shè)計發(fā)夾RNA(short-hairpin RNA,shRNA)干擾序列并構(gòu)建干擾載體,轉(zhuǎn)染胃癌HGC-27細胞。熒光實時定量PCR及蛋白質(zhì)印跡技術(shù)分別檢測轉(zhuǎn)染干擾載體后HGC-27細胞中IGF-1的表達情況。細胞增殖-毒性檢測試劑盒(cell counting kit-8,CCK-8)、劃痕實驗及Transwell侵襲實驗分別檢測下調(diào)IGF-1對體外細胞HGC-27增殖及遷移、侵襲能力的影響。結(jié)果免疫組織化學(xué)法檢測顯示,低分化胃癌組織中IGF-1陽性表達率為76%(29/38),高、中分化為53%(21/40),癌旁正常組織為27%(8/30)。不同胃癌組織與癌旁組織中IGF-1的表達差異有統(tǒng)計學(xué)意義,χ2=16.66,P0.01。并且其表達與胃癌的病理分期(χ2=7.430,P=0.007)、組織分化(χ2=4.618,P=0.032)等病理因素相關(guān)。體外不同胃癌細胞系中IGF-1均呈現(xiàn)高表達,但差異無統(tǒng)計學(xué)意義。利用干擾載體能夠有效下調(diào)胃癌細胞HGC-27中IGF-1在mRNA及蛋白質(zhì)水平的表達。與空載體轉(zhuǎn)染對照組和野生型HGC-27細胞比較,IGF-1表達下調(diào)明顯抑制胃癌細胞的體外增殖和侵襲、遷移能力。結(jié)論 IGF-1在體內(nèi)胃癌組織及體外胃癌細胞中高表達,下調(diào)IGF-1的表達能夠抑制人胃癌細胞HGC-27的增殖及體外侵襲、遷移能力。
[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.
【作者單位】： 承德醫(yī)學(xué)院附屬醫(yī)院臨床技能教學(xué)中心;承德醫(yī)學(xué)院附屬醫(yī)院病理科;承德醫(yī)學(xué)院附屬醫(yī)院老年病科;承德醫(yī)學(xué)院附屬醫(yī)院手足外科;
【分類號】：R735.2

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本文編號：1708156