Adriamycin resistance-associated prohibitin gene inhibits proliferation of human osteosarcoma MG63 cells by interacting with oncogenes and tumor suppressor genes
Adriamycin resistance-associated prohibitin gene inhibits proliferation of human osteosarcoma MG63 cells by interacting with oncogenes and tumor suppressor genes
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阿霉素耐药相关抑制基因通过与癌基因和抑癌基因相互作用抑制人骨肉瘤MG63细胞的增殖
DOI：
10.3892/ol.2016.4862
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发表时间：
2016-09-01
期刊：
影响因子：
2.9
通讯作者：
Li, Jin-Yi
中科院分区：
文献类型：
--
作者：
Du, Min-Dong;He, Kai-Yi;Li, Jin-Yi
肿瘤细胞对化疗药物的耐药性是化疗成功的主要障碍，其耐药机制尚不清楚。本研究建立了阿霉素耐药的人骨肉瘤MG-63亚系(MG-63/ADR)，并鉴定了可能与阿霉素耐药相关的差异表达蛋白。进行了双向凝胶电泳、基质辅助激光解吸电离飞行时间质谱分析和蛋白质鉴定。Western印迹分析检测MG-63/ADR细胞中的禁止素(PHB)水平。采用定量聚合酶链式反应检测阿霉素耐药相关基因。用激光扫描共聚焦显微镜观察PHB与v-myc禽髓细胞瘤病毒癌基因同源物(c-myc)、FBJ小鼠骨肉瘤病毒癌基因同源物(c-fos)、肿瘤蛋白P53和视网膜母细胞瘤1(Rb)的共定位。此外，还将人PHB的全长开放阅读框亚克隆到慢病毒载体pLVX-Puro中。并观察其对MG-63细胞的增殖率。MG-63/ADR细胞的总蛋白表达明显受到抑制。与亲本细胞相比，MG-63/ADR细胞中代表高迁移率族蛋白Box 1、RAS同源基因家族、成员A和PHB的三个显著蛋白质区域发生了显著变化。因此，PHB通过与多个癌基因或抑癌基因(c-myc、c-fos、p53和Rb)相互作用来调节MG-63/ADR细胞的化疗耐药性。此外，PHB的过表达降低了MG-63细胞的增殖率。结论：PHB是阿霉素耐药相关基因，可能通过与癌基因或抑癌基因c-myc、c-fos、p53和Rb相互作用而抑制人骨肉瘤MG-63细胞的增殖。
The resistance of cancer cells to chemotherapeutic agents is a major obstacle for successful chemotherapy, and the mechanism of chemoresistance remains unclear. The present study developed an adriamycin-resistant human osteosarcoma MG-63 sub-line (MG-63/ADR), and identified differentially expressed proteins that may be associated with adriamycin resistance. Two dimensional gel electrophoresis, matrix-assisted laser desorption ionization time-of-flight mass spectrometry analysis and a protein identification assay were performed. Western blot analysis was used to examine the prohibitin (PHB) levels in the MG-63/ADR cells. Quantitative polymerase chain reaction was utilized to detect adriamycin resistant-associated genes. Laser-scanning confocal microscope was employed to examine the colocalization of PHB with v-myc avian myelocytomatosis viral oncogene homolog (c-myc), FBJ murine osteosarcoma viral oncogene homolog (c-fos), tumor protein p53 and retinoblastoma 1 (Rb). In addition, the full length of the open reading frame of human PHB was subcloned into a lentiviral vector pLVX-puro. The proliferative rate of MG-63 cells was also investigated. The overall protein expression in MG-63/ADR cells was clearly suppressed. Three notable protein regions, representing high mobility group box 1, Ras homolog gene family, member A, and PHB, were identified to be significantly altered in MG-63/ADR cells when compared with its parental cells. Therefore, PHB modulated the chemoresistance of MG-63/ADR cells by interacting with multiple oncogenes or tumor suppressor genes (c-myc, c-fos, p53 and Rb). In addition, overexpression of PHB decreases the proliferative rate of MG-63 cells. In conclusion, PHB is an adriamycin resistance-associated gene, which may inhibit the proliferation of human osteosarcoma MG-63 cells by interacting with the oncogenes or tumor suppressor genes, c-myc, c-fos, p53 and Rb.