西妥昔单抗对A-431细胞系增殖和凋亡的影响

Effects of cetuximab on proliferation and apoptosis of A-431 cell line

  • 摘要:
      背景   皮肤鳞状细胞癌(cutaneous squamous cell carcinoma,cSCC)是临床上最常见的皮肤恶性肿瘤之一,cSCC死亡人数占所有皮肤癌死亡人数的20%。部分晚期cSCC患者无法通过手术得到有效治疗,而指南推荐这类患者采用分子靶向治疗、免疫治疗、化疗等全身系统性治疗的方式改善预后。
      目的   研究西妥昔单抗对高表达表皮生长因子受体(epidermal growth factor receptor,EGFR)的皮肤鳞状细胞癌细胞系A-431增殖和凋亡的影响。
      方法   于人类蛋白质图谱数据库中筛查并通过Western blot实验验证A-431细胞的EGFR表达情况。将A-431细胞随机分为对照组和西妥昔单抗治疗组(200 μg/mL),通过细胞增殖抑制实验(MTT法)检测西妥昔单抗对体外培养的A-431细胞增殖和毒性的影响,应用流式细胞术检测西妥昔单抗对A-431细胞凋亡的影响。
      结果   在人类蛋白质图谱数据库中对照了收录的41种正常组织及肿瘤细胞系,其中A-431细胞EGFR表达量最高。在Western blot实验中我们发现,与人永生化表皮细胞HaCat相比,A-431细胞中EGFR确为高表达状态。MTT实验表明,在200 μg/mL浓度下,西妥昔单抗对A-431细胞表现出了较强的抑制增殖作用,可使约61.35%肿瘤细胞死亡(P<0.01)。流式细胞术显示,西妥昔单抗在150 μg/mL浓度时,细胞凋亡率达到29.85%,而对照组的细胞凋亡率仅10.44%(P<0.01)。
      结论   西妥昔单抗对于高表达EGFR的皮肤鳞状细胞癌细胞A-431具有良好的肿瘤细胞杀伤能力,并可以促进肿瘤细胞的凋亡,为临床中无法接受手术治疗的皮肤鳞状细胞癌患者提供了一种治疗策略。

     

    Abstract:
      Background   Cutaneous squamous cell carcinoma (cSCC), is one of the most common skin malignant tumors in clinic, accounting for 20% of all skin cancer deaths. Some advanced cSCC patients cannot be effectively treated by operation. The guidelines recommend molecular targeted therapy, immunotherapy, chemotherapy and other systemic therapies to improve prognosis for these patients.
      Objective   To study the effect of cetuximab on the proliferation and apoptosis of cutaneous squamous cell carcinoma cell line A-431 with high expression of epidermal growth factor receptor (epidermal growth factor receptor, EGFR).
      Methods   The expression of EGFR in A-431 cells were screened from the human protein map database and verified by Western blot experiment. A-431 cells were randomly divided into control group and cetuximab treatment group (200 μ G/mL), and MTT assay was used to detect the effect of cetuximab on the proliferation and toxicity of A-431 cells in vitro. Flow cytometry was performed to detect apoptosis.
      Results   In the human protein map database, 41 kinds of normal tissues and tumor cell lines were compared, among which the expression of EGFR in A-431 cells was the highest. The expression of EGFR in A-431 cell was significantly higher than that in human immortalized epidermal cell HaCat by Western blot. MTT assay showed that cetuximab strongly inhibited the proliferation of A-431 cells at the concentration of 200 μg/mL and killed about 61.35% of tumor cells (P<0.01). Flow cytometry showed that the apoptosis rate of cetuximab at the concentration of 150 μg/mL was 29.85%, while that of the control group was only 10.44% (P<0.01).
      Conclusion   Cetuximab has good killing ability to cutaneous squamous cell carcinoma cell line A-431 with high expression of EGFR and significantly increases the apoptosis of tumor cells, which provides a therapeutic strategy for cSCC patients who cannot be treated by surgery.

     

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