CAJ | 학술논문

目的观察抑制转化生长因子-βⅡ型受体(TGF-βRⅡ)表达对食管成纤维细胞参与瘢痕形成的影响.方法利用5例食管癌术后患者吻合口狭窄处的活检组织,原代培养食管成纤维细胞并行细胞免疫化学鉴定.针对人TGF-βRⅡ基因序列设计合成的小片段RNA(siRNA)转染成纤维细胞,荧光显微镜观察转染效率；采用Western blot和逆转录-聚合酶链反应(RT-PCR)检测TGF-βRⅡ和纤维粘连蛋白的表达；细胞划痕实验评价细胞迁移能力.结果原代食管成纤维细胞培养成功,细胞抗波形蛋白免疫荧光阳性.针对TGF-βRⅡ的siRNA转染成纤维细胞48 h后转染效率达80％以上,与对照组比较明显抑制了TGF-βRⅡ和纤维粘连蛋白的表达(P＜0.05),并抑制了成纤维细胞(70.6±9.5)％的迁移能力.结论使用RNA干扰技术沉默食管成纤维细胞TGF-βRⅡ的表达抑制了其在瘢痕形成中的作用.
목적 관찰억제전화생장인자-βⅡ형수체(TGF-βRⅡ)표체대식관성섬유세포삼여반흔형성적영향.방법 이용5례식관암술후환자문합구협착처적활검조직,원대배양식관성섬유세포병행세포면역화학감정.침대인TGF-βRⅡ기인서렬설계합성적소편단RNA(siRNA)전염성섬유세포,형광현미경관찰전염효솔；채용Western blot화역전록-취합매련반응(RT-PCR)검측TGF-βRⅡ화섬유점련단백적표체；세포화흔실험평개세포천이능력.결과 원대식관성섬유세포배양성공,세포항파형단백면역형광양성.침대TGF-βRⅡ적siRNA전염성섬유세포48 h후전염효솔체80％이상,여대조조비교명현억제료TGF-βRⅡ화섬유점련단백적표체(P＜0.05),병억제료성섬유세포(70.6±9.5)％적천이능력.결론 사용RNA간우기술침묵식관성섬유세포TGF-βRⅡ적표체억제료기재반흔형성중적작용.
Objective To study the effects of inhibition of transforming growth factor-beta type Ⅱ receptor (TGF-βRⅡ ) in esophageal fibroblasts on scar formation.Methods Primary esophageal fibroblasts were cultured from endoscopic biopsy specimens of esophageal anastomotic stricture after operation in 5 patients with esophageal cancer and identified by using immunocytochemistry.TGF-βR Ⅱ specific small interfering RNA (siRNAs) designed from the human gene sequence were transfected into cultured fibroblasts.The tansfection efficiency was observed by fluorescence microscope,the expression of TGF-βRⅡand fibronectin were detected by Western blotting and reverse transcription-polymerase chain reaction ( RTPCR),and wound scratch assay was used to evaluate migration of esophageal fibroblasts.Results Primary esophageal fibroblasts were successfully cultured and positive for Vimentin.The transfection efficiency of siRNAs was above 85％ at 48 h after transfection.As compared with control groups,in TGF-βRⅡ specific siRNA targeted fibroblasts,the expression of TGF-βRⅡ and fibronectin was inhibited (P ＜ 0.05) and cell migration was decreased by (70.6 ± 9.5) ％.Conclusion TGF-βRⅡ specific siRNA targeted esophageal fibroblasts can inhibit the scar formation.