Department of Obstetrics and Gynecology, West China Second University Hospital, Sichuan University, Chengdu, Sichuan, China.
Arch Gynecol Obstet. 2011 Nov;284(5):1241-6. doi: 10.1007/s00404-010-1782-9. Epub 2010 Dec 14.
Recent evidence suggests that ADP-ribosylation factor-like tumor suppressor gene 1(ARLTS1) may act as a tumor suppressor gene. However, its role in tumor chemotherapy remains unclear. The aim of this study is to investigate the effects of ARLTS1 gene in regulation of chemosensitivity in ovarian cystadenocarcinoma cell line SKOV3.
We stably expressed wild-type (wt) ARLTS1 and empty vector (neo) in SKOV3 cells. Chemosensitivity test was carried out with four chemotherapeutic agents. Cell proliferation, cycle kinetics and apoptosis were evaluated by MTT assay and flow cytometry. Apoptosis-related proteins caspase-3 and bcl-2 were determined by Western blot analysis.
The proliferation of wtARLTS1 clones was more dramatically inhibited by all the cytotoxic agents than parental cells (P < 0.05). Increased sensitivity to chemotherapy by two to threefolds was detected in wtARLTS1 cells. The rate of apoptosis in wtARLTS1 was 60.2% treated with DDP (10× peak plasma concentration, PPC), which was dramatically higher than that of neo and parental cells (P is 0.017 and 0.020, respectively). Expression of caspase-3 and bcl-2 in parental cells declined modestly when treated with DDP, while in wtARLTS1 clones the expression of caspase-3 and bcl-2 levels declined more dramatically and become undetectable at lower DDP doses (P = 0.023 and <0.001, respectively).
Our findings suggested that ARLTS1 may facilitate chemosensitivity in ovarian cancer cells by acting synergistic with chemotherapeutic agents to induce the apoptosis signaling pathway and regulate apoptosis-related proteins.
最近的证据表明，ADP-核糖基化因子样肿瘤抑制基因 1（ARLTS1）可能作为一种肿瘤抑制基因发挥作用。然而，其在肿瘤化疗中的作用尚不清楚。本研究旨在探讨 ARLTS1 基因在调节卵巢囊腺癌 SKOV3 细胞系化疗敏感性中的作用。
我们在 SKOV3 细胞中稳定表达野生型（wt）ARLTS1 和空载体（neo）。用四种化疗药物进行化疗敏感性试验。用 MTT 法和流式细胞术检测细胞增殖、细胞周期动力学和细胞凋亡。用 Western blot 分析检测凋亡相关蛋白 caspase-3 和 bcl-2。
wtARLTS1 克隆的增殖比亲本细胞对所有细胞毒性药物的抑制更为显著（P<0.05）。wtARLTS1 细胞对化疗药物的敏感性增加了两到三倍。wtARLTS1 细胞用 DDP（10×峰值血浆浓度，PPC）处理时，凋亡率为 60.2%，明显高于 neo 和亲本细胞（P 分别为 0.017 和 0.020）。用 DDP 处理亲本细胞时，caspase-3 和 bcl-2 的表达略有下降，而在 wtARLTS1 克隆中，caspase-3 和 bcl-2 的表达下降更为显著，在较低的 DDP 剂量下无法检测到（P 分别为 0.023 和<0.001）。
我们的研究结果表明，ARLTS1 可能通过与化疗药物协同作用，诱导凋亡信号通路，调节凋亡相关蛋白，促进卵巢癌细胞的化疗敏感性。
