| RAGE在不同宫颈鳞癌细胞中的表达及意义 |
| 投稿时间：2017-06-18 修订日期：2017-08-02 点此下载全文 |
| 引用本文：周璐璐,朱雪洁,程慧慧,朱雪琼.RAGE在不同宫颈鳞癌细胞中的表达及意义[J].医学研究杂志,2018,47(4):86-89,93 |
| DOI：
10.11969/j.issn.1673-548X.2018.04.022 |
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| 基金项目:温州市科技计划项目（Y20150041） |
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| 中文摘要:目的 探讨人宫颈鳞癌细胞中晚期糖基化终末产物受体（receptor for advanced-glycation end products，RAGE）的表达水平及定位，筛选出合适的细胞用于研究RAGE对宫颈鳞癌细胞生物学行为的影响。方法 选取人宫颈鳞癌细胞SiHa、MS751、C33-A及CaSki，采用免疫细胞化学染色检测癌细胞中RAGE蛋白的定位，应用qRT-PCR法检测癌细胞中RAGE mRNA的表达水平。应用免疫印迹法检测癌细胞中RAGE蛋白的表达水平。采用酶联免疫吸附法检测4种人宫颈鳞癌细胞分别培养至12、24、36、48及72h时，上清中分泌型RAGE蛋白的表达水平。结果 RAGE蛋白主要在人宫颈鳞癌细胞的胞质和胞膜中表达，大部分位于胞质。RAGE mRNA的表达量在SiHa细胞中最高，为0.986±0.053，而在MS751细胞中的表达量相对最低，仅0.111±0.008（P<0.05）。RAGE蛋白在SiHa细胞中的表达量相对最高的（0.982±0.096），而在MS751细胞中的表达量相对最低（0.250±0.056），差异有统计学意义（P<0.05）。SiHa、MS751、C33-A及CaSki细胞各个时间点均检测到其分泌至上清中的RAGE蛋白，且4种细胞随着培养时间的延长，其分泌型RAGE的表达量逐渐增高。结论 RAGE在4种人宫颈鳞癌细胞中均有表达。C33-A和CaSki细胞适合用做后续RAGE干扰和过表达实验；SiHa细胞可在培养液中加入抗RAGE抗体改变RAGE表达量；用于后续研究RAGE对宫颈鳞癌细胞生物学行为的影响。 |
| 中文关键词:RAGE 宫颈鳞癌 SiHa C-33A CaSki MS751 |
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| Expression and Significance of the Receptor for Advanced-glycation End Products in the Different Cervical Squamous Cancer Cells |
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| Abstract:Objective To investigate the expression and localization of RAGE gene in cervical squamous cancer cell lines and to select the suitable cell line to study the effects of RAGE expression on the biological behavior of cervical squamous cell carcinomas. Methods Four different cervical squamous cancer cell lines-SiHa, MS751, C33-A and CaSki were selected. Immunocytochemistry was used to detect the localization of RAGE in cells. qRT-PCR was used to examine the expression of RAGE mRNA in cells. Western blot analysis was used to examine the expression of RAGE protein in cancer cells. Using enzyme-linked immunosorbent assay, the secreted protein of RAGE in cancer cells with different culture time (12h, 24h, 36h, 48h, 72h) was detected. Results RAGE protein was mainly expressed in the cytoplasm and membrane of human cervical squamous cancer cells, especially in cytoplasm. The expression of RAGE mRNA in SiHa was relative highest (0.986±0.053), whereas the MS751 was the lowest (0.111±0.008)(P<0.05). The expression of RAGE protein in SiHa was relative highest (0.982±0.096), whereas the MS751 was the lowest (0.250±0.056)(P<0.05). The secreted RAGE protein was detected in all four kinds of human cervical squamous cancer cells and it gradually rose with the culture time extended. Conclusion RAGE was expressed in four kinds of human cervical squamous carcinoma cells. C33-A and CaSki was suitable for further RAGE interference experiments or RAGE overexpress experiments. In addition, change the expression of RAGE through adding anti-RAGE antibody in SiHa cell culture medium. Therefore, we can choose above ways to observe the effect of RAGE on the biological behavior of cervical squamous cell carcinomas. |
| keywords:RAGE Cervical squamous cancer SiHa C-33A CaSki MS751 |
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