Role of metallothionein1h in cisplatin resistance of non-small cell lung cancer cells
Role of metallothionein1h in cisplatin resistance of non-small cell lung cancer cells
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DOI：
10.1007/s11670-009-0247-9
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发表时间：
2009-12-01
影响因子：
5.1
通讯作者：
Lu, Shi-xin
中科院分区：
文献类型：
--
作者：
Hou, Xin-fang;Fan, Qing-xia;Lu, Shi-xin
尽管铂类辅助化疗已大大改善了患者的预后，但耐药性是成功使用这种有效药物的主要障碍。金属硫蛋白（MT）在肿瘤细胞增殖、凋亡、分化、耐药性和预后等方面发挥重要作用。为探讨金属硫素1H（MT 1H）在人非小细胞肺癌（NSCLC）细胞顺铂耐药中的作用及其可能的分子机制，采用RT-PCR方法检测MT 1H在A549和A549/DDP细胞中的mRNA表达。构建了MT 1H真核表达质粒pcDNA3.1（-）-MT 1H，并转染不表达MT 1H的A549细胞。将MT 1H siRNA转染高表达MT 1H的A549/DDP细胞。RT-PCR和免疫印迹法检测MT 1H的表达。MTT法测定细胞对顺铂的敏感性。TUNEL法和流式细胞术检测细胞凋亡率。MTT法检测A549/DDP细胞中Bcl-2和Bax的表达，A549/DDP细胞中MT 1H mRNA表达阳性，而A549细胞中MT 1H mRNA表达阴性。转染MT 1H后，A549细胞MT 1H表达增强，对顺铂的敏感性降低。转染MT 1H siRNA后，A549/DDP细胞MT 1H表达降低，对顺铂敏感性增加。MT 1H siRNA干预后，顺铂诱导的A549/DDP细胞凋亡率增加，Bcl-2表达下调，Bax表达无明显变化，MT 1H过表达可促进A549细胞耐药。siRNA干扰MT 1H表达下调可有效逆转A549/DDP细胞的耐药性，其机制可能与下调Bcl-2的表达，增加顺铂诱导的细胞凋亡有关。靶向MT 1H的siRNA联合化疗可能是一种非常有前途的肺癌治疗策略。
Despite platinum-based adjuvant chemotherapy has improved greatly patients' outcomes, drug resistance poses a major impediment to the successful use of such an effective agent. Metallothioneins(MTs) are known to play putative roles in cancer cell proliferation, apoptosis, differentiation, drug resistance and prognosis. The present studiy was to investigte the role of metallethioein1H(MT1H) in cisplatin resistance of human non-small cell lung cancer(NSCLC) cell lines in vitro or its possible molecular mechanisms.MT1H mRNA expression in A549 and A549/DDP cells was detected by RT-PCR. A recombinant eukaryotic expression plasmid pcDNA3.1(-)-MT1H was constructed and transfected into A549 cells which express no MT1H. MT1H siRNA was transfected into A549/DDP cells which express MT1H highly. MT1H expression was detected by RT-PCR and Immunoblot. The chemosensitivity to cisplatin was assessed by MTT assay. Apoptosis rate was determined by Tunel and FCM. Bcl-2 and Bax were determined by immunohistochemistry.MT1H mRNA was expressed in A549/DDP but not in A549. After transfection of MT1H, MT1H expression was enhanced and the chemosensitivity to cisplatin was decreased in A549 cells. Inversely, after transfection of MT1H siRNA, MT1H expression was decreased and the chemosensitivity to cisplatin was increased in A549/DDP. The apoptosis rate induced by cisplatin was increased and Bcl-2 was down-regulated but Bax showed little change in A549/DDP cells interferred with MT1H siRNA.MT1H overexpression can promote drug resistance in A549 cells. Down-regulation of MT1H interfered with siRNA can effectively reverses the drug resistance in A549/DDP cells by down-regulating the expression of Bcl-2 and increasing cisplatin induced apoptosis. SiRNA targeting MT1H combined with chemotherapy may be a very promising strategy for treatment of lung cancer.