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目的探讨肌切蛋白(scinderin,SCIN)对胃肠道间质瘤(gastrointestinal stromal tumor,GIST)细胞增殖能力的影响。方法用shRNA慢病毒干扰技术将SCIN在GIST细胞中稳定敲低,并通过Western blot和qRT-PCR检测敲低效率。通过CCK-8实验检测伊马替尼IC50和SCIN对GIST细胞增殖能力的影响,通过平板克隆形成实验检测SCIN对GIST细胞克隆形成能力的影响,通过裸鼠皮下成瘤实验检测SCIN对移植瘤生长的影响。实验按照对GIST细胞进行的不同处理分组:WT组(野生型组,未进行转染),Mock组(阴性对照组,转染阴性对照序列),sh SCIN 1组(转染SCIN干扰序列1),sh SCIN 2组(转染SCIN干扰序列2)。结果 sh SCIN 1组和sh SCIN 2组GIST细胞在增殖能力上明显低于Mock组,增殖能力在第2~7天均有明显差异(n=4,P<0.05)。移植瘤实验中sh SCIN 1组较Mock组的移植瘤质量显著降低:sh SCIN 1组vs Mock组=(0.175 0±0.045 6)g vs(0.334 1±0.016 5)g(n=4,P<0.01)。平板克隆形成实验显示sh SCIN 1组克隆形成数显著少于Mock组,分别为59.67±5.68和123.33±2.52(n=3,P<0.01)。在使用0.57μmol/L的伊马替尼处理WT组细胞1、3、5 d后,SCIN的表达随处理时间而降低。结论敲低SCIN可以抑制GIST细胞在体外和体内环境中的增殖能力,伊马替尼可以抑制GIST细胞中SCIN的表达,SCIN可能成为GIST的治疗靶点。
Objective To investigate the effect of scindein (SCIN) on the proliferation of gastrointestinal stromal tumor (GIST) cells. Methods SCIN was stably knocked down in GIST cells by shRNA lentiviral interference technique and the knockdown efficiency was detected by Western blot and qRT-PCR. The effect of Imatinib IC50 and SCIN on the proliferation of GIST cells was detected by CCK-8 assay. The effect of SCIN on the clonality of GIST cells was detected by plate clone formation assay. The growth of transplanted tumor was detected by subcutaneous tumor formation test in nude mice Impact. The experiment was divided into groups according to different treatments on GIST cells: WT group (wild type group, untransfected), Mock group (negative control group, negative control sequence transfected), sh SCIN 1 group (transfected SCIN interference sequence 1) , sh SCIN 2 group (transfected with SCIN interference sequence 2). Results The proliferative abilities of sh SCIN 1 group and sh SCIN 2 group were significantly lower than those of Mock group (P <0.05). The proliferative ability of GIST cells differed significantly between days 2 and 7 (n = 4, P <0.05). In the xenograft experiment, the tumor mass of sh SCIN 1 group was significantly lower than that of Mock group: sh SCIN 1 vs Mock group (0.175 ± 0.045 6) g vs (0.334 ± 0.016 5) g (n = 4, P < 0.01). The results of plate clone formation assay showed that the sh SCIN 1 group had significantly fewer colonies than the Mock group, which were 59.67 ± 5.68 and 123.33 ± 2.52, respectively (n = 3, P <0.01). After treated with 0.57μmol / L imatinib for 1, 3, 5 days, the expression of SCIN decreased with the time of treatment. Conclusion Knockdown of SCIN can inhibit the proliferation of GIST cells in vitro and in vivo. Imatinib can inhibit the expression of SCIN in GIST cells, and SCIN may be the target of GIST treatment.