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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、20.0μg/mL)、CBP及OXA(终浓度均为0、2.5、5.0、10.0、20.0、40.0μg/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(IC50)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IC50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ERCC1mRNA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ERCC1mRNA和Survivin mRNA表达有关。
Objective To investigate the effects of hyperthermia combined with three generations of platinum drugs, cisplatin (DDP), carboplatin (CBP) and oxaliplatin (OXA) on the proliferation of ovarian cancer cell line SKOV3 and its possible mechanism. Methods SKOV3 cells were treated with different concentrations of DDP (0, 1.25, 2.5, 5.0, 10.0 and 20.0 μg / mL, respectively), CBP and OXA at 42 ℃ or 37 ℃ 0, 2.5, 5.0, 10.0, 20.0 and 40.0 μg / mL). The proliferation of SKOV3 cells was detected by MTT assay. The nucleotide excision and repair of SKOV3 cells were detected by qRT-PCR The expression of excision repair cross-complementing group 1 (ERCC1) and apoptosis-inhibiting gene (Survivin) were detected. Results DDP, CBP and OXA could inhibit the proliferation of ovarian cancer cell line SKOV3 in a dose-dependent manner (P <0.05). The hyperthermia at 42 ℃ enhanced the proliferation inhibition of SKOV3 cell line; (IC50) of ovarian cancer cell line OXA were (7.271 ± 0.096) μg / mL, (37.609 ± 0.779) μg / mL and (28.328 ± 0.698) μg / mL, respectively. The IC50 of platinum group were (2.075 ± 0.244) μg / mL, (19.591 ± 0.453) μg / mL and (19.089 ± 0.424) μg / mL, respectively. The sensitivities were 2.075 ± 0.244, 1.92 ± 0.044 and 1.484 ± 0.039 (P <0.05), of which the most obvious effect on DDP sensitization; in three generations of platinum drugs, hyperthermia can down-regulate ERCC1mRNA expression (P <0.05), and CBP down the most significant; hyperthermia combined with CBP, OXA The expression of Survivin mRNA was down-regulated in SKOV3 cells (P <0.05), especially in OXA group. However, the effect of DDP plus hyperthermia on Survivin mRNA expression was not statistically significant (P> 0.05). Conclusions Hyperthermia combined with three generations of platinum drugs can enhance the inhibition of proliferation of SKOV3 cells by platinum drugs and increase the sensitivity of tumor cells to platinum drugs. The mechanism of hyperthermia may be related to the down-regulation of ERCC1 mRNA and Survivin mRNA expression.