RRM1和RRM2表达水平与宫颈癌化疗的相关性研究

发布时间：2018-06-15 18:32

本文选题：核糖核苷酸还原酶亚单位M1 + 核糖核酸还原酶亚单位M2　；参考：《宁波大学》2014年硕士论文

【摘要】：目的核糖核苷酸还原酶(Ribonucleotide reductase，RR)是细胞内DNA复制和损伤修复中的限速酶，在细胞增殖以及肿瘤增殖和浸润转移中起着重要作用。宫颈癌在女性高发恶性肿瘤中位于第二位，化疗是宫颈癌治疗的一种重要手段。RR是一些抗肿瘤药物的作用靶点，吉西他滨（Gemcitabine，GEM）是目前最有效且应用最广泛的RR抑制剂。本研究通过检测宫颈癌组织和细胞中RR酶活性及亚基表达水平，探讨RR在宫颈癌发生、发展的可能作用；进一步研究吉西他滨与卡铂(Carboplatin, CBDCA)联合用药对宫颈癌细胞的作用效果。方法采用实时荧光定量PCR(qRT-PCR)、Western Blotting和CDP还原法分别检测宫颈癌组织及癌旁正常组织中RR亚基表达及酶活性，CCK-8法检测宫颈癌细胞系SiHa、CaSki对GEM、CBDCA单药及合药的敏感性，EdU法检测GEM对宫颈癌细胞DNA合成的影响，Western Blotting和免疫荧光两种方法检测药物对宫颈癌细胞的DNA损伤作用，流式细胞仪检测GEM、CBDCA单药及合药时细胞凋亡情况，并基于Chou-Talalay法计算合用指数（CI）以分析GEM与CBDCA合用对宫颈癌细胞是否具有协同作用。结果宫颈癌组织中的RRM1和RRM2mRNA水平较癌旁正常组织中的表达水平明显升高(P0.0001)，类似地，RR酶活和亚基蛋白表达水平在宫颈癌组织中的水平也上调。GEM能抑制宫颈癌细胞DNA合成，CBDCA能诱导DNA损伤，，GEM与CBDCA联合作用对宫颈癌细胞具有协同毒性作用(log10[CI]0)，GEM与CBDCA合用能促进诱导宫颈癌细胞DNA损伤产生和促进细胞凋亡率增加。结论核糖核苷酸还原酶在宫颈癌中的表达上调，细胞实验表明RR抑制剂GEM与宫颈癌一线化疗药物CBDCA合用对宫颈癌细胞产生协同毒性作用，协同作用是通过抑制DNA合成和促进细胞凋亡产生的。本研究可能为GEM联合CBDCA在宫颈癌的临床治疗应用提供实验依据。
[Abstract]:Objective ribonucleotide reductase ( RR ) is a limiting enzyme in DNA replication and repair of cervical cancer , plays an important role in cell proliferation and tumor proliferation and invasion .
To further study the effect of gemcitabine and carboplatin ( CBDCA ) on cervical cancer cells .

Methods The expression of RR subunits in cervical cancer tissues and adjacent normal tissues was detected by real - time fluorescence quantitative PCR ( qRT - PCR ) , Western Blotting and CDP reduction . The effects of GEM and CBDCA alone on DNA synthesis of cervical cancer cells were detected by the method of Western Blotting and CDP . The effect of GEM on DNA synthesis of cervical cancer cells was detected by the method of Western Blotting and Immunofluorescence .

Results The levels of RRM1 and RRM2mRNA in cervical cancer tissues were significantly higher than those in normal tissues ( P0.05 ) .

Conclusion The expression of ribonucleotide reductase in cervical cancer is up - regulated . The results suggest that the combination of the RR inhibitor GEM and the first - line chemotherapy drug CBDCA has a synergistic effect on cervical cancer cells . The synergistic effect is the inhibition of DNA synthesis and the promotion of apoptosis . This study may provide experimental basis for the application of GEM and CBDCA in the clinical treatment of cervical cancer .
【学位授予单位】：宁波大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R737.33

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