中国麻风皮肤病杂志 ›› 2017, Vol. 33 ›› Issue (7): 416-420.

• 论著 • 上一篇    下一篇

转染c-kit基因对A375细胞增殖和凋亡的影响

王正想1  张国强2  高顺强2  刘思源3    

  1. 1.河北沧州市中心医院皮肤科，河北沧州061001
    2.河北医科大学第四医院皮肤科，石家庄050010
    3.河北医科大学第三医院骨与软组织肿瘤科, 石家庄050051
  • 出版日期:2017-07-15 发布日期:2018-12-01
  • 通讯作者: 刘思源 E-mail：306734712@qq.com

The effect of c-kit on A375 cell proliferation and apoptosis

WANG Zhengxiang, ZHANG Guoqiang, GAO Shunqiang, LIU Siyuan.   

  1. 1 Institute of Dermatology, The Center Hospital of Cangzhou, Hebei 061000, China;2,Institute of Dermatology, The Fourth Hospital of Hebei Medical University, Shijiazhuang 050011,China;3 Department of Oncology in Bone and Soft Tissue Department,the Third Hospital of Hebei Medical University, Shijiazhuang 050051, China
  • Online:2017-07-15 Published:2018-12-01
  • Contact: LIU Siyuan, E-mail：306734712@qq.com

PDF (PC)

87

摘要： 目的:明确转染c-kit基因对A375细胞增殖、凋亡的影响。方法：体外培养人恶性黑素瘤A375细胞，分为A组（实验组）、B组（转染空病毒载体的阴性对照组）、C组（空白对照组）。转染后采用RT-PCR检测c-kit mRNA表达情况，在倒置相差显微镜观察A375细胞分化及形态，MTT比色分析法检测对A375细胞增殖抑制的影响，流式细胞分析术，Anexin-ⅴ-PI双染色检测细胞早期凋亡的变化。结果：转染后A组细胞出现不同程度的皱缩、破裂，漂浮细胞增多。RT-PCR半定量分析结果显示，A组扩增出c-kit cDNA 230bp大小片段，而B、C组则没有扩增出相应大小的片段。转染后Ａ组OD值低于B、C组（P＜0.05），A组早期凋亡率高于B、C组有显著性差异（P<0.05）。结论：转染c-kit后A375细胞增殖被抑制，早期凋亡增加。

关键词: 黑素瘤, 转移性, A375细胞, 早期凋亡

Abstract: Objective: To determine the effect of c-kit gene on the proliferation and apoptosis of A375 cells. Methods: Human malignant melanoma A375 cells were cultured and divided into the group A (adenovirous vector group), group B (vacant adenovirous vector group) and group C (blank control group). The expression level of c-kit mRNA was detected by RT-PCR. The differentiation and morphology of the cells were observed under the inverted microscope. The proliferation ability and apoptosis of A375 cells were detected by MTT and Annexin V-FITC/PI. Results: There was more shrinking and splinter cells in group A than those in group B and C. The c-kit cDNA 230bp was detected in group A but not in group B and C. The proliferation ability of A375 cells in group A was lower than those in group B and C, with a significant difference (P＜0.005). The apoptosis rate of A375 cells in the group A was higher than that in the group B and C (P＜0.05). Conclusion: The proliferation ability of A375 cells is inhibited and apoptosis is induced after transfected by c-kit gene.

Key words: malignant melanoma, metastasis, A375 cells, apoptosis