LY294002抑制PI3K-Akt通路对富集肝癌干细胞细胞球增殖的影响

doi:10.3760/cma.j.issn.1673-422X.2015.04.001

摘要/Abstract

摘要： 目的探讨LY294002对富集肝癌干细胞的细胞球增殖和磷脂酰肌醇3激酶蛋白激酶B（PI3KAkt）通路的影响。方法对人肝癌HepG2细胞进行无血清悬浮培养获得富集肝癌干细胞的细胞球，消化后使用LY294002（10、20、30 μmol/L）培养作为实验组，对照组不使用LY294002。细胞活性计数法检测细胞增殖活性，Western blotting检测Akt，实时定量PCR检测PI3KAkt信号通路下游基因诱骗受体3（DcR3）、哺乳动物雷帕霉素靶蛋白（mTOR）、B淋巴细胞瘤2（Bcl2）、细胞周期蛋白D1（Cyclin D1）的表达。结果与对照组相比，30 μmol/L LY294002可显著抑制富集肝癌干细胞的细胞球增殖，吸光度（A）值有显著差异［(0.14±0.03)∶(0.56±0.01), t=-8.915, P=0.000］；磷酸化Akt表达水平明显降低［(0.57±0.08)∶(0.16±0.42), t=6.027, P=0.026］；DcR3［(0.38±0.08 )∶1,t=13.060, P=0.006］、mTOR［(0.37±0.04 )∶1, t=30.363, P=0.001］、Bcl2［(0.26±0.04)∶1, t=33.554, P=0.001］、Cyclin D1［(0.1±0.02)∶1, t=63.528, P=0.000］表达明显减少。结论 LY294002可能通过抑制PI3KAkt通路而抑制富集人肝癌干细胞的细胞球的增殖。

关键词: 肝肿瘤, 肿瘤干细胞, 信号传导, 细胞球

Abstract: ObjectiveTo investigate the impact of LY294002 on the proliferation of cancer stem cellenriched spheroid cells from human hepatocellular carcinoma via regulating phosphatidylinositol3kinaseprotein kinase B (PI3KAkt) signaling pathway.MethodsThe cancer stem cellenriched spheroid cells were generated by culturing HepG2 cells in serumfree medium. LY294002 (10, 20, 30 μmol/L), an inhibitor of PI3KAkt signaling pathway, was used in the experimental groups, without used in the control group. The impact of LY294002 on the spheroid cells proliferation was confirmed by cell counting kit (CCK8 kit). The expression of Akt was tested by Western blotting. The expression of PI3KAkt signaling pathway downstream genes such as decoy receptor 3 (DcR3), mammalian target of rapamycin (mTOR), Bcell lymphoma (Bcl)2 and Cyclin D1 were tested by realtime PCR.Results30 μmol/L LY294002 could inhibit the proliferation of spheroid cells, and significant difference in the absorbance (A value) was observed between the experimental group and control group［(0.14±0.03) vs (0.56±0.01), t=-8.915, P=0.000］. The expression level of phosphorylated Akt protein increased［(0.57±0.08) vs (0.16±0.42), t=6.027, P=0.026］. The mRNA of DcR3［(0.38±0.08) vs 1, t=13.060, P=0.006］, mTOR［(0.37±0.04) vs 1, t=30.363, P=0.001］, Bcl2［(0.26±0.04) vs 1, t=33.554, P=0.001］ and Cyclin D1［(0.10±0.02) vs 1, t=63.528, P=0.000］ decreased.ConclusionLY294002 could inhibit the proliferation of cancer stem cellenriched spheroid cells from human hepatocellular carcinoma via inhibiting PI3KAkt signaling pathway.

Key words: Liver neoplasms, Neoplastic stem cells, Signal transduction, Spheroid cells

冷政伟；杨刚；李勇；石刚. LY294002抑制PI3K-Akt通路对富集肝癌干细胞细胞球增殖的影响[J]. 国际肿瘤学杂志, 2015, 42(4): 241-244.

LENG ZhengWei；Yang Gang；Li Yong；Shi-Gang. LY294002 decreases the proliferation of cancer stem cell-enriched spheroid cells from human hepatocellular carcinoma via inhibiting of PI3K-Akt signaling pathway[J]. Journal of International Oncology, 2015, 42(4): 241-244.

参考文献

［1］ Visvader JE, Lindeman GJ. Cancer stem cells: current status and evolving complexities［J］. Cell Stem Cell, 2012,10(6): 717728. ［2］ Nguyen LV, Vanner R, Dirks P, et al. Cancer stem cells: an evolving concept［J］. Nat Rev Cancer, 2012,12(2): 133143. ［3］ Leng Z, Tao K, Xia Q, et al. Kruppellike factor 4 acts as an oncogene in colon cancer stem cellenriched spheroid cells［J］. PLoS One, 2013,8(2): e56082. ［4］ Cao L, Zhou Y, Zhai B, et al. Sphereforming cell subpopulations with cancer stem cell properties in human hepatoma cell lines［J］. BMC Gastroenterol, 2011,11: 71. ［5］ Deng L, Zhang R, Tang F, et al. Ursolic acid induces U937 cells differentiation by PI3K/Akt pathway activation［J］. Chin J Nat Med, 2014,12(1): 1519. ［6］ Kantara C, O′Connell M, Sarkar S, et al. Curcumin promotes autophagic survival of a subset of colon cancer stem cells, which are ablated by DCLK1siRNA［J］. Cancer Res, 2014,74(9): 24872498. ［7］ Ponti D, Costa A, Zaffaroni N, et al. Isolation and in vitro propagation of tumorigenic breast cancer cells with stem/progenitor cell properties［J］. Cancer Res, 2005,65(13): 55065511. ［8］ Wei B, Han XY, Qi CL, et al. Coaction of spheroidderived stemlike cells and endothelial progenitor cells promotes development of colon cancer［J］. PLoS One, 2012,7(6): e39069. ［9］ Liu J, Ma L, Xu J, et al. Spheroid bodyforming cells in the human gastric cancer cell line MKN45 possess cancer stem cell properties［J］. Int J Oncol, 2013,42(2): 453459. ［10］ Yang T, Rycaj K, Liu ZM, et al. Cancer stem cells: constantly evolving and functionally heterogeneous therapeutic targets［J］. Cancer Res, 2014,74(11): 29222927. ［11］ Tomuleasa C, Soritau O, RusCiuca D, et al. Isolation and characterization of hepatic cancer cells with stemlike properties from hepatocellular carcinoma［J］. J Gastrointestin Liver Dis, 2010,19(1): 6167. ［12］ Zhu Z, Hao X, Yan M, et al. Cancer stem/progenitor cells are highly enriched in CD133+CD44+ population in hepatocellular carcinoma［J］. Int J Cancer, 2010,126(9): 20672078. ［13］ Terris B, Cavard C, Perret C. EpCAM, a new marker for cancer stem cells in hepatocellular carcinoma［J］. J Hepatol, 2010,52(2): 280281. ［14］ Zheng L, Gong W, Liang P, et al. Effects of AFPactivated PI3K/Akt signaling pathway on cell proliferation of liver cancer［J］. Tumour Biol, 2014,35(5): 40954099. ［15］姜蓬垒, 常冰梅. PI3KAkt信号通路与肿瘤化疗耐药性［J］. 国际肿瘤学杂志, 2014, 41(5)： 324327. ［16］ Jiang QG, Li TY, Liu DN, et al. PI3K/Akt pathway involving into apoptosis and invasion in human colon cancer cells LoVo［J］. Mol Biol Rep, 2014, 41(5): 33593367. ［17］ Wang XJ, Feng CW, Li M. ADAM17 mediates hypoxiainduced drug resistance in hepatocellular carcinoma cells through activation of EGFR/PI3K/Akt pathway［J］. Mol Cell Biochem, 2013,380(12): 5766.

[1]	陈红健, 张素青. 血清miR-24-3p、H2AFX与肝癌患者临床病理特征及术后复发的关系研究[J]. 国际肿瘤学杂志, 2024, 51(6): 344-349.
[2]	彭琴, 蔡玉婷, 王伟. KPNA2在肝癌中的研究进展[J]. 国际肿瘤学杂志, 2024, 51(3): 181-185.
[3]	孙国宝, 杨倩, 庄庆春, 高斌斌, 孙晓刚, 宋伟, 沙丹. 结直肠癌肝转移组织病理学生长方式研究进展[J]. 国际肿瘤学杂志, 2024, 51(2): 114-118.
[4]	向玉玲, 谭佳杰, 熊远果, 赵丽蓉, 黎晨, 张洪. 脱水淫羊藿素对肝癌细胞增殖、迁移和凋亡的影响[J]. 国际肿瘤学杂志, 2023, 50(9): 513-519.
[5]	李佳璇, 封颖璐. 糖皮质激素受体在肝癌细胞生长中的作用机制[J]. 国际肿瘤学杂志, 2023, 50(4): 241-243.
[6]	和婷, 王希, 张惠中, 刘昕阳, 王会平, 董轲. 血清TIM-3对肝癌患者诊断价值的研究[J]. 国际肿瘤学杂志, 2022, 49(9): 537-542.
[7]	张玉敏, 赵现伟, 何前进, 陈杰能. 超声造影联合血清CXCL8、CXCR2在原发性肝癌经导管动脉化疗栓塞术后疗效评估中的价值分析[J]. 国际肿瘤学杂志, 2022, 49(10): 592-596.
[8]	狄伟华, 赵雪梅. DNA损伤修复在肝癌中的研究进展[J]. 国际肿瘤学杂志, 2022, 49(10): 635-638.
[9]	杜佳航, 陈栋, 陈耀庭. 三氧化二砷抗肝癌机制及其在肝癌治疗中的研究[J]. 国际肿瘤学杂志, 2021, 48(9): 572-.
[10]	韩保俊. 组蛋白乙酰转移酶P300在肝癌组织中的表达及其临床意义[J]. 国际肿瘤学杂志, 2021, 48(7): 415-419.
[11]	魏永健, 胡进静, 李汛. CDCA8与肿瘤发生发展及干细胞干性维持的关系[J]. 国际肿瘤学杂志, 2021, 48(4): 216-219.
[12]	张玉元, 李臻, 詹鹏超, 李鑫, 叶书文, 王彩鸿, 刘杨. 肝癌标志物研究进展[J]. 国际肿瘤学杂志, 2021, 48(4): 241-245.
[13]	熊琳, 张修云, 张小余, 黎越, 徐细明. IWR-1-endo通过抑制Wnt通路影响肝癌细胞的迁移和增殖[J]. 国际肿瘤学杂志, 2021, 48(12): 711-715.
[14]	刘俊国, 张金卷, 王毅军. ALPPS中肝脏离断技术变异的临床应用进展[J]. 国际肿瘤学杂志, 2020, 47(8): 492-495.
[15]	张伟, 殷海涛, 周冲, 李向阳, 郭林. 甲磺酸阿帕替尼联合卡瑞利珠单抗治疗原发性肝癌伴肺转移一例[J]. 国际肿瘤学杂志, 2020, 47(8): 510-512.