[1]侯继院,单国用,王海霞,等.色瑞替尼对肺癌细胞H3122增殖及磷脂酰肌醇3-激酶/蛋白激酶B信号通路的影响[J].新乡医学院学报,2020,37(7):601-605.[doi:10.7683/xxyxyxb.2020.07.001]
　HOU Jiyuan,SHAN Guoyong,WANG Haixia,et al.Effect of ceritinib on proliferation of lung cancer cells H3122 and phosphatidylinositol 3-kinase/protein kinase B signaling pathway[J].Journal of Xinxiang Medical University,2020,37(7):601-605.[doi:10.7683/xxyxyxb.2020.07.001]
点击复制

色瑞替尼对肺癌细胞H3122增殖及磷脂酰肌醇3-激酶/蛋白激酶B信号通路的影响
分享到：

《新乡医学院学报》[ISSN:1004-7239/CN:41-1186/R]

卷:
37
期数:
2020年7
页码:
601-605
栏目:
基础研究
出版日期:
2020-07-05

文章信息/Info

Title:
Effect of ceritinib on proliferation of lung cancer cells H3122 and phosphatidylinositol 3-kinase/protein kinase B signaling pathway
作者:
侯继院1; 单国用1; 王海霞1; 王作培2
（1.郑州人民医院肿瘤放疗科，河南　郑州　450000；2.河南大学淮河医院心胸肿瘤外科，河南　开封　475000）
Author(s):
HOU Jiyuan1; SHAN Guoyong1; WANG Haixia1; WANG Zuopei2
(1.Department of Radiation Oncology，People′s Hospital of Zhengzhou，Zhengzhou 450000，Henan Province，China；2.Department of Cardiothoracic Oncology Surgery，Huaihe Hospital of Henan University，Kaifeng 475000,Henan Province，China)
关键词:
色瑞替尼; 肺癌细胞; 细胞增殖; 磷脂酰肌醇3-激酶/蛋白激酶B信号通路
Keywords:
ceritinib; lung cancer cell; cell proliferation; phosphatidylinositol 3-kinase/protein kinase B signal pathway
分类号:
R734.2
DOI:
10.7683/xxyxyxb.2020.07.001
文献标志码:
A
摘要:
目的　探讨色瑞替尼对肺癌细胞H3122增殖及磷脂酰肌醇3-激酶（PI3K）/蛋白激酶B（Akt）信号通路的影响。方法　将对数生长期H3122细胞随机分为空白组及低、中、高剂量色瑞替尼组，每组设5个复孔。低、中、高剂量色瑞替尼组分别加入色瑞替尼，使每孔终质量浓度分别为16.67、33.33、66.66 mg·L-1，空白组加入等量的磷酸盐缓冲液。采用甲基噻唑基四唑法检测干预24、48、72 h后各组细胞增殖抑制率,流式细胞术检测干预72 h后各组细胞凋亡率，实时荧光定量聚合酶链式反应检测干预72 h后各组细胞中PI3K、Akt、Caspase-3 mRNA相对表达量，蛋白质免疫印迹法检测干预72 h后各组细胞中PI3K、p-PI3K、Akt、p-Akt、Caspase-3蛋白表达。结果　培养24、48、72 h后，低、中、高剂量色瑞替尼组细胞增殖抑制率均高于空白组（P<0.05）,中、高剂量色瑞替尼组细胞增殖抑制率高于低剂量组（P<0.05），高剂量色瑞替尼组细胞增殖抑制率高于中剂量组（P<0.05）；低、中、高剂量色瑞替尼组细胞增殖抑制率随时间延长均呈显著上升趋势（P<0.05）。培养72 h时，低、中、高剂量色瑞替尼组细胞凋亡率均高于空白组（P<0.05），中、高剂量色瑞替尼组细胞凋亡率高于低剂量组（P<0.05），高剂量色瑞替尼组细胞凋亡率高于中剂量组（P<0.05）。4组细胞中Caspase-3 mRNA相对表达量随色瑞替尼剂量的增加呈上升趋势，组间两两比较差异均有统计学意义（P<0.05）；4组细胞中PI3K、Akt mRNA相对表达量比较差异均无统计学意义（P>0.05）。4组细胞中Caspase-3蛋白相对表达量随色瑞替尼剂量增加呈上升趋势，p-PI3K/PI3K、pAkt/Akt随色瑞替尼剂量的增加呈下降趋势，组间两两比较差异均有统计学意义（P<0.05）。结论　色瑞替尼可抑制肺癌细胞增殖，促使其凋亡，且呈一定的剂量依赖性，其作用机制可能与调控PI3K、Akt及其下游Caspase-3基因和蛋白表达有关。
Abstract:
Objective　To investigate the effect of ceritinib on the proliferation of lung cancer cells H3122 and the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt) signaling pathway.Methods　The H3122 cells in logarithmic phase were randomly divided into blank group，low-dose ceritinib group，medium-dose ceritinib group and high-dose of ceritinib group，and each group was provided with 5 replicate.The ceritinib was added to the low-,medium- and high-dose ceritinib groups so that the final concentration of each well was 16.67，33.33, 66.66 mg·L-1，and the blank group was added with the equal amount of phosphate buffer solution (PBS).The cell proliferation inhibition rate was detected by methyl thiazolyl tetrazolium method at 24，48，and 72 hours after intervention in each group.The apoptosis rate at 72 hours after intervention was detected by flow cytometry.The relative expression of PI3K，Akt，and Caspase-3 mRNA in the cells was detected by real-time fluorescent quantitative polymerase chain reaction at 72 hours after intervention.The expression of PI3K，p-PI3K，Akt，p-Akt，Caspase-3 protein in the cells was detected by Western blot at 72 hours after intervention in each group, and the p-PI3K/PI3K，p-Akt/Akt in each group were compared.Results　The cell proliferation inhibition rate in the low-，medium- and high-dose ceritinib groups was higher than that in the blank group at 24,48 and 72 hours after intervention(P<0.05).The cell prolifera tion inhibition rate in the medium- and high-dose ceritinib groups was higher than that in the low-dose ceritinib group (P<0.05).The cell proliferation inhibition rate in the high-dose ceritinib group was higher than that in the medium-dose ceritinib group (P<0.05).The cell proliferation inhibition rate in the low-, medium- and high-dose ceritinib groups increased significantly with the extension of the time(P<0.05).The apoptosis rate in the low-，medium- and high-dose groups was higher than that in the blank group (P<0.05).The apoptosis rate in the medium- and high-dose ceritinib groups was lower than that in the low-dose ceritinib group (P<0.05).The apoptosis rate in the high-dose ceritinib group was higher than that in the medium-dose ceritinib group (P<0.05).The relative expression of Caspase-3 mRNA in the four groups showed an increasing trend with the increase of the dose of ceritinib,and the pairwise comparison between the groups showed statistically significant differences (P<0.05).There was no statistic difference in the relative expression of PI3K,Akt mRNA among the four groups(P>0.05）.The relative expression of Caspase-3 protein in the four groups showed an increasing trend with the increase of the dose of ceratinib, while the p-PI3K/PI3K and pAkt/Akt showed a decreasing trend with the increase of the dose of ceratinib, and the pairwise comparison between the groups showed statistically significant differences (P<0.05).Conclusion　Ceritinib can inhibit the proliferation of lung cancer cells and promote their apoptosis, and it is dose-dependent.Its mechanism may be related to regulating the expressions of PI3K, Akt and the downstream Caspase-3 gene and protein.

参考文献/References:

［1］　柴文杰， 王良友， 乔冬菊， 等.2010～2016年浙江省台州市肺癌发病和死亡趋势分析［J］.中国肿瘤，2018，33(7):509-513.
［2］　HIRSCH F R，SCAGLIOTTI G V，MULSHINE J L，et al.Lung cancer:current therapies and new targeted treatments［J］.Lancet，2017，389(10066):299-311.
［3］　GADGEEL S M.Sequencing of ALK inhibitors in ALK+ non-small cell lung cancer［J］.Curr Treat Options Oncol，2017，18(6):36.
［4］　ZHOU Z Y，MUTEBI A，HAN S，et al.Cost-effectiveness of ceritinib in previously untreated anaplastic lymphoma kinase-positive metastatic non-small cell lung cancer in the United States［J］.J Med Econ，2018，21(6):577-586.
［5］　SOUSA V M L，CARVALHO L.Heterogeneity in lung cancer ［J］.Pathobiology，2018，85(1/2):96-107.
［6］　DURUISSEAUX M，ESTELLER M.Lung cancer epigenetics:from knowledge to applications ［J］.Semin Cancer Biol，2018，51:116-128.
［7］　LI G，DAI W R，SHAO F C.Effect of ALK-inhibitors in the treatment of non-small cell lung cancer:a systematic review and meta-analysis ［J］.Eur Rev Med Pharmacol Sci，2017，21(15):3496-3503.
［8］　SOLOMON B J，KIM D W，WU Y L，et al.Final overall survival analysis from a study comparing first-line crizotinib versus chemotherapy in ALK-mutation-positive non-small-cell Lung cancer ［J］.J Clin Oncol，2018，36(22):2251-2258.
［9］　SPAGNUOLO A，MAIONE P，GRIDELLI C.Evolution in the treatment landscape of non-small cell lung cancer with ALK gene alterations:from the first- to third-generation of ALK inhibitors［J］.Expert Opin Emerg Drugs，2018，23(3):231-241.
［10］　胡鑫， 张会云， 边南南， 等.第二代ALK抑制剂治疗ALK+非小细胞肺癌有效性单组率的Meta分析［J］.中国循证医学杂志，2018，18(9):918-923.
［11］　ERP A E M V，HILLEBRANDT-ROEFFEN M H S，HOUDT L V，et al.Targeting anaplastic lymphoma kinase (ALK) in rhabdomyosarcoma (RMS) with the second-generation ALK inhibitor ceritinib［J］.Target Oncol，2017，12(6):815-826.
［12］　JANOSTIAK R,MALVI P,WAJAPEYEE N.Anaplastic lymphoma kinase confers resistance to BRAF kinase inhibitors in melanoma［J］.Science,2019,16:453-467.

更新日期/Last Update: 2020-07-05