本文選題：重組質(zhì)粒　切入點：AIM-　出處：《腫瘤防治研究》2015年11期 　論文類型：期刊論文

【摘要】：目的探討pCMV AIM-1 antisence重組質(zhì)粒對卵巢癌裸鼠移植瘤的體內(nèi)抑瘤效應。方法(1)人卵巢癌細胞A2780轉(zhuǎn)染pCMV空載體和pCMV AIM-1 antisense重組質(zhì)粒,G418篩選出抗性克隆,擴增后產(chǎn)生的穩(wěn)定轉(zhuǎn)染株用于實驗;(2)通過RT-PCR、Western blot檢測未轉(zhuǎn)染A2780細胞(對照組)、A2780轉(zhuǎn)染pCMV空載體和pCMV AIM-1 antisense重組質(zhì)粒三組AIM-1 m RNA和蛋白的表達;(3)將穩(wěn)定轉(zhuǎn)染的pCMV空載體和pCMV AIM-1 antisense重組質(zhì)粒的A2780細胞注入裸鼠皮下,建立裸鼠移植瘤模型,未轉(zhuǎn)染A2780細胞注入裸鼠皮下作為對照,觀察三組裸鼠移植瘤生長情況,計算腫瘤重量和腫瘤體積,并進行統(tǒng)計對比分析;(4)SP免疫組織化學法檢測移植瘤組織切片AIM-1的表達。結(jié)果 (1)A2780對照組和轉(zhuǎn)染pCMV空載體組AIM-1 m RNA和蛋白表達無明顯改變;而A2780轉(zhuǎn)染pCMV AIM-1 antisense重組質(zhì)粒組細胞AIM-1 m RNA和蛋白表達明顯下降;(2)A2780對照組、A2780轉(zhuǎn)染pCMV空載體組和A2780轉(zhuǎn)染pCMV AIM-1 antisense重組質(zhì)粒組的裸鼠腫瘤重量分別為(1.93±0.21)g、(1.90±0.39)g和(0.77±0.34)g;移植瘤體積分別為(1 011.7±235.6)mm3、(981.2±162.3)mm3和(468.7±119.6)mm3,重組質(zhì)粒組與空載體組和對照組相比,差異有統(tǒng)計學意義(P0.01);pCMV AIM-1 antisense重組質(zhì)粒的抑瘤率達53.8%。(3)SP免疫組織化學法檢測瘤體組織切片AIM-1的表達情況,見A2780對照組和轉(zhuǎn)染pCMV空載體組AIM-1表達沒有明顯改變,而A2780轉(zhuǎn)染pCMV AIM-1antisense重組質(zhì)粒組AIM-1的表達明顯減少。重組質(zhì)粒組AIM-1蛋白表達陽性率和陽性標記指數(shù)均明顯低于對照組和空載體組(P0.01)。結(jié)論 pCMV AIM-1 antisense重組質(zhì)粒對A2780移植瘤生長有明顯抑制作用。
[Abstract]:Objective to investigate the inhibitory effect of pCMV AIM-1 antisence recombinant plasmid on ovarian cancer xenografts in nude mice. Methods: human ovarian cancer cell line A2780 was transfected with pCMV empty vector and pCMV AIM-1 antisense recombinant plasmid G418 was used to screen the resistant clones. The stable transfection strain was used in the experiment. The untransfected A2780 cell line (the control group was transfected with pCMV empty vector and pCMV AIM-1 antisense recombinant plasmid three groups AIM-1 m RNA and protein expression) was detected by RT-PCRG Western blot. A2780 cells of pCMV AIM-1 antisense recombinant plasmid were injected subcutaneously into nude mice. The transplanted tumor model of nude mice was established, and the untransfected A2780 cells were injected subcutaneously into nude mice as control. The growth of transplanted tumor was observed in three groups, and the tumor weight and volume were calculated. The expression of AIM-1 m RNA and protein in transplanted tumor sections were detected by immunohistochemical method. Results the expression of AIM-1 m RNA and protein in the control group and the empty pCMV vector group were not significantly changed. However, the expression of AIM-1 m RNA and protein in A2780 transfected pCMV AIM-1 antisense recombinant plasmid group was significantly decreased. The tumor weight of Nude mice transfected with pCMV empty vector and pCMV AIM-1 antisense recombinant plasmid was 1.93 鹵0.21 鹵0.39 g and 0.77 鹵0.34 g, respectively, and the tumor volume of transplanted tumor was 0.77 鹵0.34 g, respectively, and the tumor weight of nude mice transfected with A2780 was 1.93 鹵0.21 鹵0.39 g and 0.77 鹵0.34 g respectively, and that of A2780 transfected with pCMV AIM-1 antisense was 1.90 鹵0.39 g and 0.77 鹵0.34 g respectively. Compared with empty vector group and control group, 1011.7 鹵235.6 mm ~ (-3) and 468.7 鹵119.6 mm ~ (-3) mm3 and 468.7 鹵119.6 mm ~ (-3) mm3 respectively, the recombinant plasmid group was compared with the empty vector group and the control group. The inhibitory rate of pCMV AIM-1 antisense recombinant plasmid was 53.8%. The expression of AIM-1 in tumor sections was detected by immunohistochemistry. There was no significant change in AIM-1 expression in A2780 control group and empty pCMV vector transfection group. The positive rate and labeling index of AIM-1 protein expression in the recombinant plasmid group were significantly lower than those in the control group and empty vector group (P 0.01). Conclusion the expression of pCMV AIM-1antisense recombinant plasmid in A2780 transplanted tumor was significantly lower than that in the control group and empty vector group. Growth has obvious inhibitory effect.
【作者單位】： 武漢市中心醫(yī)院中西醫(yī)結(jié)合腫瘤科;武漢市中心醫(yī)院影像診斷科;
【分類號】：R737.31

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