【摘要】：目的:構(gòu)建G蛋白信號調(diào)節(jié)蛋白2(GPSM2)穩(wěn)定高表達的胰腺癌細胞株,探討GPSM2與人胰腺癌細胞遷移能力的關(guān)系。方法:構(gòu)建GPSM2基因過表達質(zhì)粒(pCMV-Tag3B-GPSM2)并鑒定,將人胰腺癌MIA-PaCa-2細胞分別轉(zhuǎn)染pCMV-Tag3B-GPSM2(GPSM2轉(zhuǎn)染組)或p CMV-Tag3B空載體(陰性對照組),以無處理的MIA-Pa Ca-2細胞為空白對照,用RT-PCR檢測各組細胞GPSM2 mRNA表達;Westernblot檢測各組細胞GPSM2、β-連環(huán)蛋白(β-catenin)的表達;用Transwell實驗檢測各組胰腺癌細胞遷移能力。結(jié)果:成功構(gòu)建了GPSM2穩(wěn)定高表達的重組細胞株。與空白對照組比較,GPSM2轉(zhuǎn)染組細胞GPSM2 mRNA表達量明顯上調(diào),達前者73.3倍、GPSM2、β-catenin蛋白表達量明顯升高、遷移細胞計數(shù)明顯增加(均P0.05)。此外,胰腺癌細胞中GPSM2與β-catenin的表達水平呈明顯的正向線性關(guān)系(P0.05)。陰性對照組與空白對照組間各指標的差異均無統(tǒng)計學意義(均P0.05)。結(jié)論:上調(diào)胰腺癌細胞中GPSM2的表達能增加胰腺癌細胞的遷移能力,該作用可能與β-catenin蛋白表達升高有關(guān)。
[Abstract]:Aim: to construct a stable and high expression of G protein signal regulated protein 2 (GPSM2) pancreatic cancer cell line and to explore the relationship between GPSM2 and migration ability of human pancreatic cancer cells. Methods: GPSM2 gene overexpression plasmids (pCMV-Tag3B-GPSM2) were constructed and identified. MIA-PaCa-2 cells were transfected into pCMV-Tag3B-GPSM2 (GPSM2 transfection group) or pCMV-Tag3B empty vector (negative control group) respectively. Untreated MIA-Pa Ca-2 cells were used as blank control. The expression of GPSM2, 尾 -catenin (尾 -catenin) was detected by RT-PCR and the migration ability of pancreatic cancer cells was detected by Transwell assay. Results: the recombinant cell line with stable and high expression of GPSM2 was successfully constructed. Compared with the control group, the expression of GPSM2 mRNA and 尾 -catenin protein in the GPSM2 transfected group was 73.3 times higher than that in the control group, and the number of migration cells was significantly increased (P0.05). In addition, the expression of GPSM2 and 尾 -catenin in pancreatic cancer cells showed a positive linear relationship (P0.05). There was no significant difference between the negative control group and the blank control group (P0.05). Conclusion: upregulating the expression of GPSM2 in pancreatic cancer cells can increase the migration ability of pancreatic cancer cells, which may be related to the increased expression of 尾 -catenin protein.
【作者單位】： 江蘇大學附屬醫(yī)院普通外科;
【基金】：江蘇省自然科學基金資助項目(BK2012704) 江蘇省博士后科研計劃基金資助項目(1302096B)
【分類號】：R735.9

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本文編號：2218450