Luo Si, Pan Zhen, Liu Shuang, Yuan Shujing, Yan Nianlong
Department of Biochemistry and Molecular Biology, School of Basic Medical Science, Nanchang University, Nanchang, Jiangxi 330006, P.R. China.
Oncol Lett. 2018 Jan;15(1):483-488. doi: 10.3892/ol.2017.7309. Epub 2017 Oct 31.
Hepatoblastoma (HB) is the most type of common pediatric liver cancer. The primary chemotherapy drug for HB is cisplatin (DDP). However, patients readily develop intrinsic and acquired resistance, and severe side effects to treatment. Sphingomyelin synthase 2 (SMS2) is a key enzyme involved in the generation of sphingomyelin (SM), which is able to regulate cell proliferation, apoptosis and differentiation. The death receptors (DRs) have important functions in DDP-induced apoptosis. However, whether SMS2 is able to modulate cell apoptosis through the DR signaling pathway remains unknown. To investigate this question, SMS2 was overexpressed in HepG2 cells and treated with 3.5 mg/l cisplatin in the present study. After 24 h, the expression of SMS2, avian myelocytomatosis viral oncogene homolog (c-Myc), DR4, DR5 and caspase-3 was analyzed. Furthermore, cell viability was quantified, and apoptosis was assessed by western blot and flow cytometry analysis as well as Cell Counting kit-8. The results of the present study revealed that overexpression of SMS2 was able to increase the expression of c-Myc, cleaved caspase-3, DR4 and DR5 compared with the control group (P<0.05, n=3), and increase the levels of apoptosis in the SMS2 + DDP group, compared with the control (P<0.001, n=3). These results indicate that overexpression of SMS2 is able to improve sensitivity of HepG2 cells to DDP by increasing the expression of c-Myc, DR4 and DR5 in HepG2 cells. This increased sensitivity may decrease intrinsic and acquired resistance of chemotherapy in HB, and reduce the associated severe side effects in pediatric patients.
肝母细胞瘤(HB)是最常见的小儿肝癌类型。HB的主要化疗药物是顺铂(DDP)。然而,患者很容易产生内在性和获得性耐药,并且治疗会产生严重的副作用。鞘磷脂合酶2(SMS2)是参与鞘磷脂(SM)生成的关键酶,能够调节细胞增殖、凋亡和分化。死亡受体(DRs)在DDP诱导的凋亡中具有重要作用。然而,SMS2是否能够通过DR信号通路调节细胞凋亡仍不清楚。为了研究这个问题,本研究在HepG2细胞中过表达SMS2并用3.5mg/l顺铂进行处理。24小时后,分析SMS2、禽成髓细胞瘤病毒癌基因同源物(c-Myc)、DR4、DR5和半胱天冬酶-3的表达。此外,对细胞活力进行定量,并通过蛋白质免疫印迹、流式细胞术分析以及细胞计数试剂盒-8评估细胞凋亡。本研究结果显示,与对照组相比,SMS2过表达能够增加c-Myc、裂解的半胱天冬酶-3、DR4和DR5的表达(P<;0.05,n=3),并且与对照组相比,SMS2+DDP组的细胞凋亡水平增加(P<;0.001,n=3)。这些结果表明,SMS2过表达能够通过增加HepG2细胞中c-Myc、DR4和DR5的表达来提高HepG2细胞对DDP的敏感性。这种增加了的敏感性可能会降低HB化疗的内在性和获得性耐药,并减少小儿患者相关的严重副作用。