摘要
目的研究胃癌细胞中KIAA0101蛋白的表达,并探讨其表达下调对胃癌MKN-45细胞增殖、细胞周期和侵袭力的影响。方法利用Western blot检测3株胃癌细胞系(MKN-28、SGC-7901和MKN-45)中KIAA0101蛋白的表达。将KIAA0101小干扰RNA(siRNA)和对照siRNA分别转染胃癌MKN-45细胞,利用Cell Counting Kit-8(CCK-8试剂盒)检测细胞增殖的变化,利用流式细胞术检测细胞周期分布的改变,最后采用Boyden小室检测细胞侵袭力的变化。结果胃癌MKN-45细胞中KIAA0101蛋白的相对表达水平显著高于MKN-28和SGC-7901细胞,3组之间差异有统计学意义(P〈0.05)。CCK-8结果显示,与未处理组和对照siRNA组相比,KIAA0101 siRNA组中胃癌MKN-45细胞的增殖受到明显抑制(P〈0.05)。细胞周期分析结果显示,KIAA0101 siRNA组[(61.47±0.89)%]在G0/G1期的细胞数比率显著高于未处理组[(47.43±0.85)%]和对照siRNA组[(48.43±0.73)%;F=271.653,P=0.000]。进一步Boyden小室体外侵袭实验结果显示,KIAA0101 siRNA组(61.51±4.76)中MKN-45细胞穿过Matrigel的细胞数显著低于未处理组(138.74±10.16)和对照siRNA组(132.93±11.25;F=65.949,P=0.000)。结论KIAA0101表达下调能明显抑制胃癌细胞的增殖、诱导细胞周期静止和降低细胞的侵袭能力,因而有望成为胃癌治疗的新靶点。
Objective To investigate the expression of KIAA0101 protein in gastric carcinoma cells, and to explore the effects of its down-regulation on the cell proliferation, cell cycle and invasion. Methods Western blot was used to detect KIAA0101 protein expression in three gastric carcinoma cell lines including MKN-28, SGC-7901 and MKN-45. KIAA0101 siRNA and control siRNA were utilized to transfect MKN-45 cells, respectively. CCK-8 was used to analyze the changes of cell proliferation, and flow cytometry to examine the changes of cell cycle distribution. Finally, Boyden chamber was used to detect the ability of cell invasion. Results Relative level of KIAA0101 protein in MKN-45 cells was significantly higher than those in MKN-28 and SGC-7901 cells, and there was significant difference among the three cell lines (P 〈0. 05 ). The result of CCK-8 study demonstrated that, compared with untreated group and control siRNA group, the proliferation of MKN-45 cells in KIAA0101 siRNA group was significantly inhibited (P 〈 0. 05). Additionally, the result of cell cycle analysis revealed that the percentage of cell number in G0/G1 phase in KIAA0101 siRNA group [ ( 61.47 ± 0. 89 ) % ] was significantly higher than those in untreated group [(47.43 ±0.85)%] and control siRNA group [(48.43 ±0.73)%;F = 271.653,P =0.000]. Further, Boyden chamber assay showed that the cell numbers migrated to Matrigel in KIAA0101 siRNA group (61.51 ±4. 76) were significantly lower than those in untreated group ( 138.74 ± 10.16) and control siRNA group (132.93 ± 11.25;F =65.949, P =0.000). Conclusions Down- regulation of KIAA0101 expression leads to an inhibition of cell proliferation, cell cycle and cell invasion. It may provide a novel target for the treatment of patients with gastric carcinoma.
出处
《中华病理学杂志》
CAS
CSCD
北大核心
2012年第8期553-557,共5页
Chinese Journal of Pathology
基金
河南省教育厅科技攻关项目(2009B320008)
河南省卫生厅科技攻关课题(200804055)
关键词
胃肿瘤
RNA干扰
细胞周期
细胞系
肿瘤
Stomach neoplasms
RNA interference
Cell Cycle
Cell line, tumor