本文關(guān)鍵詞： PROK RNA干擾 結(jié)直腸癌 細胞增殖 遷移　出處：《基因組學與應用生物學》2017年04期 　論文類型：期刊論文

【摘要】：為研究前動力蛋白1(PROK1)對結(jié)直腸癌細胞增殖與遷移的作用機制,設(shè)計合成3條針對PROK1基因的靶向干擾序列,采用熒光顯微鏡觀察干擾片段的轉(zhuǎn)染情況,實時熒光定量PCR分析靶向干擾PROK1基因后PROK1 mRNA下調(diào)的表達水平,采用MTT、平板克隆形成試驗、transwell遷移實驗檢測敲除PROK1后對HT29細胞增殖和遷移的影響。結(jié)果顯示:熒光標記的Cy3-siRNA成功轉(zhuǎn)染到HT29細胞中;3條靶向PROK1基因siRNA-62、siRNA-341、siRNA-1121轉(zhuǎn)染HT29細胞48 h后,沉默效率分別為35%、70%、50%;與陰性對照組相比,siRNA-341組細胞增殖明顯被抑制,24 h抑制最顯著(p0.01);siRNA-341組的細胞克隆形成率(19.3±1.989 9)與空脂質(zhì)體對照組(30.3±3.780 6)和陰性對照組(32.3±2.404 1)相比顯著減少(p0.05);siRNA-341組通過小室遷移的細胞數(shù)(76±9.539 3)與空脂質(zhì)體對照組(212±2.645 7)和陰性對照組(193±7.539 8)相比明顯減少(p0.01)。本研究發(fā)現(xiàn)PROK1基因促進結(jié)直腸癌細胞的增殖和轉(zhuǎn)移,PROK1基因有望成為治療結(jié)直腸癌潛在的靶位點。
[Abstract]:In order to study the mechanism of Prok1) on the proliferation and migration of colorectal cancer cells, three targeted interference sequences targeting PROK1 gene were designed and synthesized. The transfection of interference fragments was observed by fluorescence microscope. Real-time fluorescence quantitative PCR was used to analyze the down-regulation of PROK1 mRNA expression after targeted interference with PROK1 gene. The effects of PROK1 knockout on the proliferation and migration of HT29 cells were detected by using MTT and plate clone formation assay. The results showed that three PROK1 targeting siRNA-62siRNA-341siRNA-1121 were successfully transfected into HT29 cells for 48 h. Compared with the negative control group, the cell proliferation of the siRNA-341 group was significantly inhibited for 24 h, and the cell clone formation rate was significantly decreased in the p0.01 siRNA-341 group (19.3 鹵1.989 9) compared with the empty liposome control group (30.3 鹵3.786) and the negative control group (32.3 鹵2.404 1). Compared with empty liposome control group (212 鹵2.645 7) and negative control group (193 鹵7.539 8), the number of cells migrating through the cell of p0.05siRNA-341 group decreased significantly (p 0.01). It was found that PROK1 gene could promote the proliferation and metastasis of colorectal cancer cell line PROK1 gene. A potential target for colorectal cancer.
【作者單位】： 中南大學湘雅醫(yī)學院醫(yī)學檢驗系;湖南省腫瘤醫(yī)院暨中南大學湘雅醫(yī)學院附屬腫瘤醫(yī)院;
【基金】：湖南省長沙市科技計劃項目(k1501016-31) 長沙市科技項目(K1403158-61) 中南大學國家級大學生創(chuàng)新類項目(201610533027)共同資助
【分類號】：R735.34

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