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热疗联合三代铂类药物对卵巢癌细胞株SKOV3的影响

Effects of Hyperthermia Combined Platinum-based Drugs on Ovarian Cancer Cell Lines SKOV3

  • 摘要: 目的 探讨热疗分别联合三代铂类药物即顺铂(DDP)、卡铂(CBP)、奥沙利铂(OXA)对卵巢癌细胞株SKOV3增殖的影响及其可能机制。方法 SKOV3细胞采用热疗(42 ℃)或常温(37 ℃)联合不同浓度的DDP(终浓度分别为0、1.25、2.5、5.0、10.0、20.0 μg/mL)、CBP及OXA(终浓度均为0、2.5、5.0、10.0、20.0、40.0 μg/mL)处理, MTT法检测对SKOV3细胞增殖的影响,采用实时荧光定量PCR(qRT-PCR)技术检测SKOV3细胞经相应处理后核苷酸切除修复交叉互补组基因(excision repair cross-complementing group 1, ERCC1)、凋亡抑制基因(Survivin)的表达变化。结果 DDP、CBP、OXA对卵巢癌细胞株SKOV3的增殖呈抑制作用,并呈剂量依赖性(P<0.05),42 ℃热疗能增强铂类药物对SKOV3细胞株的增殖抑制作用; DDP、CBP、OXA对卵巢癌细胞株增殖的半数抑制浓度(IC50)分别为(7.271±0.096) μg/mL、(37.609±0.779)μg/mL、(28.328±0.698) μg/mL,42 ℃热疗联合上述铂类药物后IC50分别为(2.075±0.244) μg/mL、(19.591±0.453) μg/mL、(19.089±0.424) μg/mL,增敏倍数分别为2.075±0.244、1.92±0.044、1.484±0.039(P<0.05),其中对DDP的增敏效果最显著;在三代铂类药物中,热疗均能下调ERCC1 mRNA的表达(P<0.05),且CBP下调最显著;热疗联合CBP、OXA作用于SKOV3时均能下调Survivin mRNA的表达(P<0.05),其中OXA组更显著,但DDP联合热疗对Survivin mRNA表达的影响差异无统计学意义(P>0.05)。结论 热疗联合三代铂类药物能增强铂类药物对SKOV3细胞的增殖抑制作用,增加肿瘤细胞对铂类药物的敏感性,其增敏机制可能与下调ERCC1 mRNA和Survivin mRNA表达有关。

     

    Abstract: Objective To investigate the effects and mechanisms of hyperthermia combined with various platinum-based drugs cis-platinum (DDP), carboplatin (CBP), oxaliplatin (OXA) on the proliferation and apoptosis of ovarian cancer cell lines SKOV3. Methods SKOV3 cells were treated with different concentrations of anticancer drugs DDP (final concentration respectively 0, 1.25, 2.5, 5.0, 10.0, 20.0 μg/mL), CBP and OXA (both final concentration respectively 0, 2.5, 5.0, 10.0, 20.0, 40 μg/mL) at a temperature of 42 ℃ for hyperthermia or 37 ℃ for normal temperature. Methyl thiazolyl tetrazolium (MTT) method was used to test growth ratios of ovarian cancer cell lines SKOV3. Real-time PCR was adopted to detect the expression level of excision repair cross-complementing group 1 (ERCC1) and Survivin mRNA in SKOV3 cells. Results DDP, CBP and OXA inhibited the growth of SKOV3 in a dose-dependent manner (P<0.05). Hyperthermia could increase the sensitivity of SKOV3 to cis-platinum,carboplatin and oxaliplatin (P<0.05). The half inhibitory concentration (IC50) values of DDP, CBP and OXA were (7.271±0.096) μg/mL, (37.609±0.779) μg/mL and (28.328±0.698) μg/mL respectively. When combined with hyperthermia, the IC50 values of DDP, CBP, and OXA were (2.075±0.244) μg/mL, (19.591±0.453) μg/mL, (19.089±0.424) μg/mL (P<0.05). The increased sensitivity index was 2.075±0.244 for cis-platinum, 1.92±0.044 for carboplatin, 1.484±0.039 for oxaliplatin. After the treatment of hyperthermia, the expression of ERCC1 and Survivin mRNA showed downward trend. ERCC1 decreased more significantly in the group of hyperthermia combined with carboplatin, and Survivin decreased more significantly in the group of hyperthermia combined with oxaliplatin (P<0.05). Conclusion Hyperthermia can enhance the sensitivity of ovarian cancer SKOV3 cells to platinum-based drugs, which may be related to the down regulation of ERCC1 and Survivin expression.

     

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