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- 2017
SPHK-1在非小细胞肺癌H460细胞耐药株中的作用
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Abstract:
摘要：目的 探讨鞘氨醇激酶-1(SPHK-1)/1-磷酸鞘氨醇(S1P)、NFκB p65信号通路在非小细胞肺癌(NSCLC)耐药细胞中发挥其耐药性的作用。方法 建立肺癌耐药细胞株H460/DDP并鉴定其生物学特性，并通过Western blot、RT-PCR实验检测并比较耐药株和亲本细胞中的SPHK-l、S1P及NFκB相关信号蛋白的表达情况。结果 成功建立了肺癌耐药细胞株H460/DDP，并对其进行了耐药性评估(IC50H460/DDP=50.62μg/mL，RIH460/DDP=2.95)；在药物浓度为10～80μg/mL时，肺癌耐药细胞株H460/DDP的细胞存活率高于肺癌细胞株H460，其差异具有统计学意义(P<0.01)；肺癌H460耐药细胞中SPHK-1、S1P及NFκB p65表达水平较亲本细胞明显增高，差异具有统计学意义(P=0.0415、P=0.0465、P=0.0218，P<0.05)；且耐药细胞核内NFκB p65蛋白表达也显著增高。结论 SPHK-1/S1P、NFκB p65信号通路在肺癌H460耐药细胞中对发挥其耐药性具有重要作用。
ABSTRACT: Objective To examine the role of SPHK-1/S1P and NFκB p65 signal pathway in non-small cell lung cancer (NSCLC) drug-resistant cells. Methods The drug-resistant cell line of lung cancer H460/DDP was constructed and its biological characteristics were identified successfully. The expression of SPHK-l was tested by RT-PCR and Western blot methods. S1P and some proteins related to NFκB pathway were studied by Western blot. Results The drug-resistant lung cancer cell line H460/DDP was constructed and its drug-resistant ability was evaluated (IC50H460/DDP=50.62μg/mL, RIH460/DDP=2.95). Cisplatin at a concentration of 10-80μg/mL significantly decreased cell death of drug-resistant cell line (P<0.01). Western blot assay analysis showed that overexpressions of SPHK-l, S1P and NFκB p65 were significantly higher in drug-resistant cell line than in their parent cell line H460 (P=0.0415, P=0.0465, P=0.0218). RT-PCR method revealed that SPHK-1 mRNA was overexpressed in drug-resistant cell line compared with that in their parent cell line H460 (P<0.05). More NFκB p65 protein in cell nucleus was expressed in drug-resistant cells than in parent cells. Conclusion SPHK-1/S1P and NFκB p65 signal pathway may play an improtant role in the drug-resistant H460 cell line in non-small cell lung cancer
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