PDECGF特异性RNA抑制片段诱导裸鼠膀胱癌细胞凋亡的研究.doc

PDECGF特异性RNA抑制片段诱导裸鼠膀胱癌细胞凋亡的研究
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作者:金宁张灵肖博晗常喜华
【摘要】目的利用RNA干扰技术在裸鼠膀胱癌动物模型中通过抑制靶向沉默血小板源性内皮生长因子基因(PDECGF)的表达,探讨其在活体动物肿瘤组织中的作用。方法将psiPDECGF1及2稳定转染的阳性T24细胞株及其他对照组直接接种于裸鼠皮下,观察肿瘤细胞生长及成瘤情况、测定肿瘤体积的变化;免疫组化检查PDECGF蛋白表达;TUNEL法检测肿瘤细胞凋亡。结果裸鼠体内成瘤实验发现siRNA2组第21天时成瘤率只有50%,而其他各组成瘤率为100%。siRNA2组生长速度明显慢于其他3组,siRNA2组不仅成瘤延迟,且生长速度明显慢于其他3组(P<)。免疫组化证实siRNA2组肿瘤组织PDECGF 蛋白表达最低; TUNEL检测siRNA2组细胞凋亡数明显多于其他3组(P<)。结论 PDECGF siRNA载体稳定转染细胞株体内实验可见膀胱癌细胞成瘤率降低,成瘤延迟及肿瘤生长速度减慢,癌细胞凋亡增多。PDECGF siRNA在裸鼠体内依然对膀胱癌细胞有明显抑制作用。
【关键词】膀胱癌;PDECGF;RNA干扰;凋亡
【Abstract】 Objective To research the PDECGF gene
′s function on the animal tumor by inhibiting the expression of the PDECGF in the nude mice bladder carcinoma animal model via siRNA The bladder carcinoma nude mice model was established by the stable transfected bladder carcinoma cells to observe tumor’s growth and detect the tumor cells apoptosis by TUNEL kit which can assay sliced DNA in tumor The form rate of tumor was only 50% in 21st day after inoculation of T24 cell in siRNA2 group through bearing tumor experiment in vivo. The rate of form tumor was 100% in other groups. The growth velocity was slower in RNAi2 group than that of other groups(P<). The expression of PDECGF protein was lowest in siRNA2 group. The cell apoptosis counts in siRNA2 group was most than that of other group