肿瘤研究与临床
腫瘤研究與臨床
종류연구여림상
CANCER RESEARCH AND CLINIC
2013年
9期
581-584,587
,共5页
尹宝慧%贾秀红%李建厂%李有杰
尹寶慧%賈秀紅%李建廠%李有傑
윤보혜%가수홍%리건엄%리유걸
基因,同源盒%RNA干扰%肺腺癌LETP-a-2细胞
基因,同源盒%RNA榦擾%肺腺癌LETP-a-2細胞
기인,동원합%RNA간우%폐선암LETP-a-2세포
Genes,homeobox%RNA interference%Lung cancer LETP-a-2 cells
目的 体外设计、合成及筛选高效沉默同源盒基因A7 (HOXA7)表达的小干扰RNA(siRNA),并研究沉默HOXA7对人类肺腺癌细胞株LETP-a-2增殖、凋亡的影响.方法 设计、合成三对针对HOXA7基因的siRNA(siRNA1、siRNA2、siRNA3)和一对阴性对照(siRNAnc),分别用阳离子脂质体介导转染LETP-a-2细胞.实验分6组:细胞对照组、脂质体对照组、siRNAnc组、siRNA1组、siRNA2组、siRNA 3组,反转录聚合酶链反应(RT-PCR)和Western blot分别检测各组HOXA7 mRNA和蛋白表达情况,四甲基偶氮唑蓝(MTT)比色法和流式细胞术分别检测沉默HOXA7后LETP-a-2细胞增殖、凋亡情况.结果 siRNA成功转染LETP-a-2细胞,三对siRNA均能有效沉默HOXA7表达,其中以siRNA2沉默HOXA7的效果最佳,在mRNA和蛋白水平沉默率分别为(57.344±4.743)％和(52.219±0.550)％；三对siRNA沉默HOXA7后均能抑制LETP-a-2细胞增殖并促进其凋亡,其中以siRNA2组效果最明显,48 h细胞增殖抑制率可达(48.144±4.992)％,凋亡率达(26.613±0.612)％.结论 筛选出高效沉默HOXA7的siRNA2能有效抑制LETP-a-2细胞增殖并促进其凋亡,提示HOXA7可能成为肿瘤基因治疗的新靶点.
目的 體外設計、閤成及篩選高效沉默同源盒基因A7 (HOXA7)錶達的小榦擾RNA(siRNA),併研究沉默HOXA7對人類肺腺癌細胞株LETP-a-2增殖、凋亡的影響.方法 設計、閤成三對針對HOXA7基因的siRNA(siRNA1、siRNA2、siRNA3)和一對陰性對照(siRNAnc),分彆用暘離子脂質體介導轉染LETP-a-2細胞.實驗分6組:細胞對照組、脂質體對照組、siRNAnc組、siRNA1組、siRNA2組、siRNA 3組,反轉錄聚閤酶鏈反應(RT-PCR)和Western blot分彆檢測各組HOXA7 mRNA和蛋白錶達情況,四甲基偶氮唑藍(MTT)比色法和流式細胞術分彆檢測沉默HOXA7後LETP-a-2細胞增殖、凋亡情況.結果 siRNA成功轉染LETP-a-2細胞,三對siRNA均能有效沉默HOXA7錶達,其中以siRNA2沉默HOXA7的效果最佳,在mRNA和蛋白水平沉默率分彆為(57.344±4.743)％和(52.219±0.550)％；三對siRNA沉默HOXA7後均能抑製LETP-a-2細胞增殖併促進其凋亡,其中以siRNA2組效果最明顯,48 h細胞增殖抑製率可達(48.144±4.992)％,凋亡率達(26.613±0.612)％.結論 篩選齣高效沉默HOXA7的siRNA2能有效抑製LETP-a-2細胞增殖併促進其凋亡,提示HOXA7可能成為腫瘤基因治療的新靶點.
목적 체외설계、합성급사선고효침묵동원합기인A7 (HOXA7)표체적소간우RNA(siRNA),병연구침묵HOXA7대인류폐선암세포주LETP-a-2증식、조망적영향.방법 설계、합성삼대침대HOXA7기인적siRNA(siRNA1、siRNA2、siRNA3)화일대음성대조(siRNAnc),분별용양리자지질체개도전염LETP-a-2세포.실험분6조:세포대조조、지질체대조조、siRNAnc조、siRNA1조、siRNA2조、siRNA 3조,반전록취합매련반응(RT-PCR)화Western blot분별검측각조HOXA7 mRNA화단백표체정황,사갑기우담서람(MTT)비색법화류식세포술분별검측침묵HOXA7후LETP-a-2세포증식、조망정황.결과 siRNA성공전염LETP-a-2세포,삼대siRNA균능유효침묵HOXA7표체,기중이siRNA2침묵HOXA7적효과최가,재mRNA화단백수평침묵솔분별위(57.344±4.743)％화(52.219±0.550)％；삼대siRNA침묵HOXA7후균능억제LETP-a-2세포증식병촉진기조망,기중이siRNA2조효과최명현,48 h세포증식억제솔가체(48.144±4.992)％,조망솔체(26.613±0.612)％.결론 사선출고효침묵HOXA7적siRNA2능유효억제LETP-a-2세포증식병촉진기조망,제시HOXA7가능성위종류기인치료적신파점.
Objective To design and screen small interefere RNA (siRNA) targeting of HOXA7,and to investigate the effect of the siRNA on human lung cancer LETP-a-2 cells proliferation and apoptosis in vitro.Methods Three pairs of siRNA targeting of HOXA7 and one pair of siRNA for negative control were transfected respectively into LETP-a-2 cells through cationic liposome.The mRNA and proteion expression levels of HOXA7 were observed by reverse transcription polymerase chain reaction (RT-PCR) and Western blot analysis.The effect of HOXA7 siRNA on growth and apoptosis of LETP-a-2 cells were measured by MTT and flow cytometry.Results All the three pairs of siRNA could inhibit HOXA7 expression effectively,among which siRNA2 got the best effects,the silence rates were (57.344±4.743) ％ on mRNA level and (52.219±0.550) ％ on protein leval.The proliferation was inhibited and the apoptosis was promoted by the siRNA targeting HOXA7 in LETP-a-2 cells,among which siRNA2 got the favourite results,the inhibitory rate was (48.144±4.992) ％ and the apoptosis rate was (26.613±0.612) ％.Conclusion The siRNA2 targeting of HOXA7 enrolls in promoting apoptosis and inhibiting grows of LETP-a-2 cells,indicating that manipulation of HOXA7 expression may be a potential therapeutic strategy for cancer.
