摘要
背景与目的:表皮生长因子受体在多种上皮源性肿瘤中过表达,与肿瘤的发生、发展密切相关,是肿瘤治疗的重要靶点。本研究采用针对EGFR的小干扰RNA(siRNA),探讨其在不同类型肿瘤细胞A431、HeLa、SPC-A-1中的RNA干扰效应。方法:化学合成针对EGFR的siRNA,转染A431、HeLa、SPC-A-1细胞,通过定量RT-PCR、免疫荧光染色和流式细胞仪检测EGFR表达;通过集落形成实验观察细胞集落形成能力,分析不同类型肿瘤细胞的RNA干扰效应。结果:转染siRNA-EGFR后,3种肿瘤细胞的EGFR表达均明显下调。在A431细胞,其mRNA水平下调73.9%,蛋白水平下调77.0%。在HeLa和SPC-A-1细胞,mRNA水平的下调分别为44.6%和57.7%,蛋白水平下调61.3%和65.2%。EGFR下调后细胞集落形成能力均出现抑制,A431、HeLa和SPC-A-1的集落形成抑制率分别为27.2%、53.9%和59.1%。结论:siRNA-EGFR可在不同类型肿瘤细胞中产生RNA干扰效应,抑制细胞集落形成能力。RNA干扰技术在开发广谱抗肿瘤靶向治疗药物中具有应用价值。
Background and purpose： The epidermal growth factor receptor （EGFR） is commonly overexpressed in a variety of solid tumors, and has important roles in cancer pathogenesis and progression. EGFR thus provides a rational target for cancer therapy. We studied siRNA-mediated inhibition of epidermal growth factor receptor expression and its biologic effects in different human cancer cell lines （A431, HeLa and SPC-A-1）. Methods： Cells were transfected with chemically synthesized siRNA-EGFR. EGFR mRNA was quantified by real-time PCR and was detected by immunofluorescence staining and flow cytometry. The biologic effects on cell growth were assessed by colony-formation assay. Results： siRNA-EGFR sig- nificantly decreased mRNA level of EGFR by 73.9 %, 44.6 % and 57.7 %, protein expression of EGFR by 77.0 %, 61.3 % and 65.2 %, and reduced colony number by 27.2 %, 53.9 % and 59.1% in A431, HeLa and SPC-A-1, respectively. Conclusions： Our data suggested that RNA interference could downregulate EGFR and inhibit colony forming ability and EGFR expression at mRNA/protein levels in human cancer cell lines with different pathological types, siRNA could be one of the promising strategies in future targeted cancer therapy.
出处
《中国癌症杂志》
CAS
CSCD
2007年第4期278-282,共5页
China Oncology
基金
上海市科委重大项目(04DZ19208)。
关键词
表皮生长因子受体
RNA干扰
肿瘤靶向治疗
epidermal growth factor receptor
RNA interference
targeted cancer therapy
