杜仲总多糖EOP高调肺癌LLC细胞中caspase3表达来抑制肿瘤细胞增殖的机制研究
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摘要
目的研究杜仲总多糖EOP通过激活caspase3途径诱导细胞凋亡来抑制肿瘤细胞增殖的机制。方法首先用MTT法选择EOP和CDDP抑制LLC细胞和KMB-17细胞的最佳作用浓度和时间,然后用最佳浓度的EOP和CDDP作用于LLC细胞,采用细胞划痕实验,检测EOP和CDDP对LLC细胞迁移的影响。凋亡细胞/坏死细胞鉴别试剂盒:最后提取细胞的总RNA和蛋白质,分别检测caspase3的mRNA水平和蛋白水平的表达。结果 EOP和CDDP抑制LLC细胞和KMB-17细胞增殖的最佳作用浓度为4.0μg/ml,最佳作用时间为48 h。细胞划痕实验结果显示,EOP和CDDP均可显著抑制肿瘤细胞LLC的侵袭迁移,划痕细胞覆盖率降低。RT-PCR和western blot结果显示,EOP组的caspase3的mRNA和蛋白的表达水平均显著高于对照组。结论杜仲总多糖(EOP)可抑制肺癌细胞生长,是通过激活caspase3途径来诱导肺癌细胞凋亡,为杜仲总多糖应用于临床提一定的理论供据。
Objective To study the mechanism of total polysaccharides EOP induced tumor cells apoptosis by activating caspase3 approach to inhibit tumor cell proliferation. Methods First,identify the optimum concentration and time of EOP and CDDP challenge,cell activity was respectively detected by MTT after treating lung cancer cell line LLC and KMB-17. Then use the best concentration of EOP and CDDP role in LLC cells,testing EOP and CDDP affect LLC cell migration. Apoptotic cells/Dead cells identification kit:Finally cells extracted total RNA and protein,respectively detect caspase3 mRNA level and the expression of protein levels. Results The best concentration was 4. 0 ug/ml,optimal duration was 48 h of EOP and CDDP inhibit LLC cells and KMB 17 cell proliferation. Cell scratch experimental results showed that the EOP and CDDP can significantly inhibit tumor cell LLC,migration,Nick cell coverage declines. RT-PCR and western blot,according to the results of EOP group caspase3 mRNA level and the expression of protein levels were significantly higher than the control group. Conclusion Extraction of total polysaccharides( EOP) can inhibit the growth of lung cancer cells,induced lung cancer cells apoptosis by activated caspase3 way. Providing a theory for application of eucommia bark total polysaccharides to clinical.
Objective To study the mechanism of total polysaccharides EOP induced tumor cells apoptosis by activating caspase3 approach to inhibit tumor cell proliferation. Methods First,identify the optimum concentration and time of EOP and CDDP challenge,cell activity was respectively detected by MTT after treating lung cancer cell line LLC and KMB-17. Then use the best concentration of EOP and CDDP role in LLC cells,testing EOP and CDDP affect LLC cell migration. Apoptotic cells/Dead cells identification kit:Finally cells extracted total RNA and protein,respectively detect caspase3 mRNA level and the expression of protein levels. Results The best concentration was 4. 0 ug/ml,optimal duration was 48 h of EOP and CDDP inhibit LLC cells and KMB 17 cell proliferation. Cell scratch experimental results showed that the EOP and CDDP can significantly inhibit tumor cell LLC,migration,Nick cell coverage declines. RT-PCR and western blot,according to the results of EOP group caspase3 mRNA level and the expression of protein levels were significantly higher than the control group. Conclusion Extraction of total polysaccharides( EOP) can inhibit the growth of lung cancer cells,induced lung cancer cells apoptosis by activated caspase3 way. Providing a theory for application of eucommia bark total polysaccharides to clinical.
引文
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[10]龚传明,屈磊,胡海涛,等.非小细胞肺癌Bim和caspase3蛋白表达检测及临床意义[J].临床军医杂志,2010,38(2):256-258.
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[12]王靖华,陈龙邦,黄文斌,等.survivin和caspase-3在非小细胞肺癌中的表达及其与肿瘤细胞凋亡的关系[J].中国肺癌杂志,2005,8(5):435-439.
[2]辛晓明,王大伟,赵娟,等.杜仲总多糖抗肿瘤作用的实验研究[J].医药导报,2009,28(6):719-721.
[3]徐贤柱,饶华,蔡险峰,等.杜仲叶多糖对小鼠免疫功能影响的研究[J].时珍国医国药,2013,24(3):541-542.
[4]苏卓,郭诚,梁韬.杜仲多糖对链脲佐菌素致糖尿病小鼠的作用[J].中国实验方剂学杂志,2016,22(14):159-162.
[5]刘华,闫立萍,王小军.红芪多糖诱导肺腺癌细胞凋亡与Bax/Bcl-2表达的研究[J].西北国防医学杂志,2016,37(4):211-213.
[6]王强,杨志雄,廖思.Fas/Fas L和caspase3在肺癌中的表达及其临床意义[J].现代肿瘤医学,2009,17(2):248-251.
[7]高俊,陈洪雷.caspase3/PARP蛋白在肺癌组织中的表达及其临床意义[J].数理医药学杂志,2006,19(1):34-36.
[8]王晓红,赵玲,郭莉,等.survivin和caspase3在非小细胞肺癌中的表达及相关性研究[J].现代生物医学进展,2006,6(9):8-11.
[9]吕志强,张惠忠,李海刚,等.非小细胞肺癌FHIT和caspase3蛋白的表达及临床意义[J].中山大学学报医学科学版,2005,26(s1):134-136.
[10]龚传明,屈磊,胡海涛,等.非小细胞肺癌Bim和caspase3蛋白表达检测及临床意义[J].临床军医杂志,2010,38(2):256-258.
[11]刘莹,王静,吴逸明,等.MGMT、CASPASE3在非小细胞肺癌中的表达及相关性研究[J]临床医学.2010,26(3):71-73.
[12]王靖华,陈龙邦,黄文斌,等.survivin和caspase-3在非小细胞肺癌中的表达及其与肿瘤细胞凋亡的关系[J].中国肺癌杂志,2005,8(5):435-439.