河北医科大学学报

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应用RNA干扰技术沉默人恶性黑素瘤细胞株A375中PPO基因的实验研究

  

  1. 1.河北省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,河北 石家庄 050011;
    3.河北医科大学第三医院骨与软组织肿瘤科,河北 石家庄 050051
  • 出版日期:2017-07-25 发布日期:2017-07-05
  • 作者简介:王正想(1982-),男,河北沧州人,河北省沧州市中心医院主治医师,医学硕士,从事皮肤疾病诊治研究。
  • 基金资助:
    国家自然科学基金资助项目(30440086)

Experimental study on PPO gene silencing in human malignant melanoma cell line A375 by RNA interference#br#

  1. 1.Department of Dermatology, Cangzhou Center Hospital, Hebei Province, Cangzhou 061000, China;
    2.Department of Dermatology, the Fourth Hospital of Hebei Medical University, Shijiazhuang
    050011, China; 3.Department of Bone Oncology, the Third Hospital of Hebei
    Medical University, Shijiazhuang 050051, China
  • Online:2017-07-25 Published:2017-07-05

摘要: [摘要]〓
〖HTH〗目的〖HTSS〗〖KG*2〗探讨RNA干扰技术沉默PPO基因对人恶性黑素瘤细胞株A375细胞生长的影响。
〖HTH〗方法〖HTSS〗〖KG*2〗用PPO腺病毒载体转染A375细胞,采用RTPCR方法检测A375细胞PPO的表达变化,采用MTT比色分析法检测对A375细胞增殖的影响,采用流式细胞术,Annexin ⅤFITC/PI双染色检测A375细胞早期凋亡的变化。
〖HTH〗结果〖HTSS〗〖KG*2〗实验组人黑色素瘤A375细胞PPO基因 mRNA校正值低于空载体组和阴性对照组(P<005)。转染PPO基因后3组第7天时细胞增殖OD值高于第1、3、5天,第5天时细胞增殖OD值高于第1、3天,第3天时细胞增殖OD值高于第1天(P<005);转染PPO基因后第1、3、5、7天时实验组细胞增殖OD值低于空载体组和阴性对照组(P<005)。转染PPO基因后3组第7天凋亡率高于第1、3、5天,第5天凋亡率高于第1、3天,第3天凋亡率高于第1天(P<005);第1、3、5、7天时实验组凋亡率高于空载体组和阴性对照组(P<005)。
〖HTH〗结论〖HTSS〗〖KG*2〗沉默PPO基因能诱导A375细胞早期凋亡,从而抑制A375细胞生长增殖。

关键词: 黑色素瘤, 基因沉默, RNA干扰

Abstract: [Abstract]  Objective〖HTSS〗〓To investigate the effect of silencing PPO gene on the growth of human malignant melanoma cell line A375 by RNA interference.
〖HTH〗〖WTHZ〗Methods〖HTSS〗〓 A375 cells were transfected with PPO adenovirus vector, expression of PPO was detectded by RTPCR in the A375 cell, MTT colorimetric analysis method was used to detect the effects on the proliferation of A375 cells, by flow cytometry and Annexin ⅤFITC/PI double staining, changes of early apoptosis were detected.
〖HTH〗〖WTHZ〗Results〖HTSS〗〓 The expression of PPO mRNA was decreased in the transfection group than in the nontransfection group and negative virus group(P<0.05). After transfected with PPO, the OD values of the three groups at 7 d were higher than that at 1 d, 3 d and 5 d, the OD values of cells at 5 d were higher than at 1 d and 3 d, the OD values of cells at 3 d were higher than at 1 d(P<0.05). The OD value of transfection group was higher than the nontransfection group and negative virus group(P<005). After transfected, the apoptosis rate of the three groups at 7 d was higher than that at 1 d, 3 d and 5 d, the apoptosis rate of cells at 5 d was higher than that at 1 d and 3 d, the apoptosis rate of cells at 3 d was higher than that at 1 d(P<0.05). The apoptosis rate of transfection group was higher than the nontransfection group and negative virus group(P<005).
〖HTH〗〖WTHZ〗Conclusion〖HTSS〗〓PPO gene silence can inhibit the proliferation and induce early apoptosis in A375 cells.

Key words: melanoma, gene silencing, RNA interference