本文選題：宮頸癌 + hTERT基因　； 參考：《中國老年學雜志》2017年17期

【摘要】：目的研究siRNA靶向沉默h TERT基因后對宮頸癌Caski細胞特性的影響。方法設計、合成特異性針對h TERT mRNA的siRNA,將其克隆入p Genesil-1.1質粒中,重組成h TERT mRNA-siRNA的表達載體,導致目的基因沉默,是由電轉染入宮頸癌Caski細胞,從而產(chǎn)生RNA干擾作用。應用Western印跡法及RT-PCR法檢測沉默h TERT基因后對于宮頸癌Caski細胞的蛋白及mRNA的表達情況;檢測細胞周期運用流式細胞儀;沉默h TERT基因后宮頸癌Caski細胞的增殖能力可通過CCK-8增殖實驗檢測。沉默h TERT基因后宮頸癌Caski細胞的侵襲能力可通過小室侵襲實驗檢測。結果 h TERT基因沉默48 h以后,與空白組及未轉染組相比,轉染組宮頸癌Caski細胞的蛋白及mRNA表達出現(xiàn)明顯降低(P0.05);細胞周期的檢測結果顯示S期的細胞數(shù)目明顯減低,轉染組的宮頸癌Caski細胞停留于G0/G1周期,空白組、轉染組及未轉染組相比差異顯著(P0.05);宮頸癌Caski細胞在轉染組的增殖過程中被明顯抑制(P0.05);轉染組穿過濾膜的細胞數(shù)量明顯減少(P0.05)。結論 Si RNA靶向沉默h TERT基因電轉染后,使宮頸癌Caski細胞的增殖和遷移能力得到抑制,一定程度上改善了宮頸癌患者的預后。
[Abstract]:Objective to study the effect of hTERT gene silencing by siRNA on the characteristics of cervical cancer Caski cells. Methods siRNAs specifically targeting hTERT mRNA were synthesized and cloned into pGenesil-1.1 plasmid, which was recombined into the expression vector of hTERT mRNA-siRNA, resulting in the silencing of the target gene, which was induced by electrotransfection into cervical cancer Caski cells, resulting in RNA interference. Western blot and RT-PCR were used to detect the expression of protein and mRNA in human cervical cancer Caski cells after hTERT gene silencing, and flow cytometry was used to detect the cell cycle. The proliferative ability of cervical cancer Caski cells after hTERT gene silencing can be detected by CCK-8 proliferative assay. The invasive ability of cervical cancer Caski cells after hTERT gene silencing can be detected by chamber invasion assay. Results after 48 h of hTERT gene silencing, the expression of protein and mRNA of cervical cancer Caski cells in transfected group was significantly lower than that in blank group and untransfected group (P0.05), and the cell cycle detection showed that the number of S phase cells was significantly decreased. In the transfected group, the number of cervical cancer Caski cells was significantly inhibited in the transfected group (P0.05), while in the blank group, the transfected group and the untransfected group were significantly inhibited (P0.05); the number of cells passing through the filter membrane was significantly decreased in the transfected group (P0.05). Conclusion Si RNA targeted silencing hTERT gene can inhibit the proliferation and migration of cervical cancer Caski cells, and improve the prognosis of cervical cancer patients to some extent.
【作者單位】： 天津市靜海縣醫(yī)院婦產(chǎn)科;天津醫(yī)科大學總醫(yī)院;
【分類號】：R737.33

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