SPAG9/MKK3/p38 axis is a novel therapeutic target for liver cancer
SPAG9/MKK3/p38 axis is a novel therapeutic target for liver cancer
复制标题
DOI：
10.3892/or.2019.6987
复制
发表时间：
2019-04-01
期刊：
影响因子：
4.2
通讯作者：
Fu, Yin
中科院分区：
文献类型：
--
作者：
Luo, Shudi;Ren, Biqiong;Fu, Yin
精子相关抗原9（SPAG 9）是几种癌症的生物标志物和潜在的治疗靶点;然而，其与肝癌进展的关系尚不清楚。本研究的目的是确定SPAG 9是否调节肝癌的增殖。应用免疫组化和细胞免疫荧光技术检测SPAG 9在人肝癌组织和肝癌细胞HepG 2中的表达和定位。用脂质体Lipofectamine（TM）2000将针对SPAG 9的小干扰RNA（siRNA）瞬时转染HepG 2细胞，CCK-8法和流式细胞仪检测细胞增殖、凋亡和细胞周期进程; Western blotting检测SPAG 9、JNK、p38、MKK 3和MKK 6的表达，免疫共沉淀法检测SPAG 9与JNK的相互作用。20例肝癌患者中有16例（80%）SPAG 9过表达。该蛋白定位于从患者获得的肝癌细胞和HepG 2细胞的细胞质和细胞核中。缺失SPAG 9可抑制HepG 2细胞的增殖，促进细胞凋亡，并使细胞周期阻滞于S期。此外，与未用靶向SPAG 9的siRNA处理的HepG 2细胞相比，SPAG 9缺陷的细胞具有降低的JNK、p38和MKK 3表达。在本研究中，发现SPAG 9通过SPAG 9/MKK 3/p38轴调节肝癌细胞的细胞增殖、凋亡和细胞周期进程。该轴是肝癌治疗的新靶点。
Sperm-associated antigen 9 (SPAG9) is a biomarker and potential therapeutic target for several cancers; however, its involvement in liver cancer progression is not clear. The aim of the present study was to determine whether SPAG9 regulates proliferation of liver cancer. Immunohistochemistry and cell immunofluorescence were used to confirm the expression and the localization of SPAG9 in human liver cancer tissues and the liver cancer-derived HepG2 cells. A small interfering RNA (siRNA) designed to target SPAG9 was transiently transfected into HepG2 cells using Lipofectamine (TM) 2000, and proliferation, apoptosis and cell cycle progression were analyzed using CCK-8 assay and flow cytometry; western blotting was used to detect the expression of SPAG9, JNK, p38, MKK3 and MKK6, and co-immunoprecipitation was used to assess the interaction between SPAG9 and JNK. SPAG9 was overexpressed in 16 out of 20 (80%) patients with liver cancer. The protein was localized in both the cytoplasm and nucleus of liver cancer cells obtained from patients and in HepG2 cells. Depletion of SPAG9 inhibited the proliferation of HepG2 cells, promoted apoptosis and arrested the cell cycle at the S phase. Moreover, cells deficient in SPAG9 had decreased expression of JNK, p38 and MKK3 compared to HepG2 cells not treated with an siRNA targeting SPAG9. In the present study, SPAG9 was revealed to regulate cell proliferation, apoptosis and cell cycle progression in liver cancer cells through the SPAG9/MKK3/p38 axis. This axis is a novel therapeutic target for liver cancer.