美洲大蠊提取物诱导Hct 116细胞凋亡的实验研究
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摘要
目的:探讨美洲大蠊提取物诱导人结肠癌Hct 116细胞凋亡的作用。方法:用不同浓度的美洲大蠊提取物7.0、7.5、8.0、8.5、9.0mg/ml处理人结肠癌Hct 116细胞,采用MTS法检测细胞增殖的变化并计算IC_(50)值;HE染色和Hoechst33258荧光染色观察细胞形态学变化;TUNEL染色和Annexin V-FITC/PI双标记检测Hct 116细胞凋亡;PI/RNase流式检测Hct 116细胞周期变化。结果:与对照组相比,美洲大蠊提取物7.0、7.5、8.0、8.5、9.0mg/ml处理Hct 116细胞24h具有增殖抑制的作用,抑制率为15.99%～51.55%且具有浓度依赖性,24h的IC_(50)值为9.05mg/ml;HE染色显示细胞出现凋亡形态学变化,Hoechst33258荧光染色、TUNEL阳性染色、Annexin V+/PI-单阳性和Annexin V+/PI+双阳性显示细胞凋亡增多且与美洲大蠊提取物作用浓度呈正相关。美洲大蠊提取物7.5mg/ml可阻滞Hct 116细胞于G0/G1期。结论:PKA可抑制Hct 116细胞增殖,具有诱导Hct 116细胞凋亡的作用。
Objective: To investigate the apoptosis of Hct116 induced by Periplaneta americana extract( PKA). Methods: HCT116 cells were treated with PKA( 7. 0、7. 5、8. 0、8. 5、9. 0 mg/ml),the cell proliferation rate was measured by MTS. The morphological changes of Hct116 cells were observed by HE and Hoechast33258 staining. The apoptosis was detected by TUNEL and Annexin V-FITC/PI,and the cell cycle was analyzed by PI/RNase Flow Cytometry. Results: Compared with the control group,PKA( 7. 0、7. 5、8. 0、8. 5、9. 0 mg/ml) had the inhibitory effect on HCT116 cells proliferation,the inhibition rate was 15. 99% ～ 51. 55%,with concentration-effect relationship. The IC50 of PKA was 9. 053 mg/ml. HE staining showed apoptotic morphological changes. The results of Hoechst33258、TUNEL + 、Annexin V +/PI-和Annexin V +/PI + showed the increase of apoptosis and had the concentration-effect relationship to PKA. Hct116 cell cycle was delayed in G0/G1 Phase by 7. 5 mg/ml PKA. Conclusion: PKA can inhibit the proliferation and induce the apoptosis of Hct116.
Objective: To investigate the apoptosis of Hct116 induced by Periplaneta americana extract( PKA). Methods: HCT116 cells were treated with PKA( 7. 0、7. 5、8. 0、8. 5、9. 0 mg/ml),the cell proliferation rate was measured by MTS. The morphological changes of Hct116 cells were observed by HE and Hoechast33258 staining. The apoptosis was detected by TUNEL and Annexin V-FITC/PI,and the cell cycle was analyzed by PI/RNase Flow Cytometry. Results: Compared with the control group,PKA( 7. 0、7. 5、8. 0、8. 5、9. 0 mg/ml) had the inhibitory effect on HCT116 cells proliferation,the inhibition rate was 15. 99% ～ 51. 55%,with concentration-effect relationship. The IC50 of PKA was 9. 053 mg/ml. HE staining showed apoptotic morphological changes. The results of Hoechst33258、TUNEL + 、Annexin V +/PI-和Annexin V +/PI + showed the increase of apoptosis and had the concentration-effect relationship to PKA. Hct116 cell cycle was delayed in G0/G1 Phase by 7. 5 mg/ml PKA. Conclusion: PKA can inhibit the proliferation and induce the apoptosis of Hct116.
引文
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[20]朱小勤,陈珮,黄中迪,等.中药调控血液肿瘤细胞周期的研究进展.上海中医药杂志,2015,49(9)∶83～86.
[2]Liu J,Zhou L,He L,et al.Periplaneta Americana extract may attenuate renal fibrosis through inhiting Janus Tyrosine 2/signal transducer and activator of transcriptio 3 pathway. Pharmacogy.2018,102∶1～8.
[3]Qin Song,Qiheng Gou,Yuxin Xie,et al.Periplaneta americana extracts promote skin wound healing via nuclear factor kappa B canonical pathway and extracellular signal-regulated kinase signaling.Evidence-Based Complementary and Alternative Medicine,2017,1～12.
[4]Qin Song,Yunxin Xie,Qiheng Gou,et al.JAK/STAT3 and Smad3 activities requrired for the wound healing properties of Perplaneta americana extrats.2017, 40(2)∶465～473.
[5]常旭,王聪,欧红利,等.美洲大蠊多肽对MFC荷瘤小鼠免疫影响的初步探索.免疫学杂志,2017,33(7)∶564～569.
[6]唐霞光,张春妹,刘嘉,等.美洲大蠊不同提取物清除自由基及抗脂质过氧化活性研究.食品工业科技,2016,37(13)∶83～85,91.
[7]董京千.美洲大蠊抗肿瘤作用研究进展.中国中医药现代远程教育.2017,15(6)∶145～147.
[8]曾娟妮,杨敏,刘筱,等.美洲大蠊提取液调控大鼠难愈合创面微丝、桥粒实验研究.中国民族民间医药,2018,27(2)∶45～47.
[9]张菌,魏永凯,李玥,等.美洲大蠊提取物Ento-D对醋酸诱导的急性溃疡性结肠炎大鼠的作用及机制研究.中华中医杂志,2018,33(2)∶304～308.
[10]李奇娟,王战国,刘巧,等.美洲大蠊研究现状及其研究中关键问题分析与展望.中国中药杂志,2018,43(7)∶1507～1516.
[11]陈万青,孙可欣,郑荣寿,等.2014年中国分地区恶性肿瘤发病和死亡分析.癌症监测, 2018,27(1)∶1～14.
[12]何正春,王晓雨,杨雷香,等.美洲大蠊提取物对3株消化系统肿瘤细胞的细胞毒性研究.中国药业,2009,18(9)∶11～12.
[13]余昕,周洁, 欧丽兰,等.美洲大蠊不同提取物及不同提取方法对肿瘤细胞的细胞毒性作用.中国实验方剂学杂志,2016,22(15)∶153～156.
[14]张丹,朱伟,余昕,等.美洲大蠊多肽提取物对SMMC-7721细胞凋亡及Bcl-2和Bax蛋白表达的影响.中国医药药学杂志,2017,37(4)∶328～332.
[15]邹俊波,熊永爱.康复新液与顺铂、伊立替康联用对大鼠癌HCT 116荷瘤小鼠肿瘤生长和造血系统的影响.世纪科学技术-中医药现代化杂志,2016,18(5)∶854～862.
[16]沈佳雯,罗会,宁倪,等.近年中药治疗结肠癌的药理作用研究进展.中华中医药学刊,2017,35(10)∶2544～2546.
[17]赵相轩,温锋,任莹,等.细胞凋亡在结肠癌治疗中作用的研究进展.肿瘤学杂志,2016,22(11)∶946～950.
[18]Klener P Jr,Andera L,Klener P,et al.Cell signaling pathways in the pathogenesis and therapy of haematologic malignancies∶overview of therapeutic approches.Folia Biol,2006,52(4)∶ 119～136.
[19]丁大伟,章永红.以细胞凋亡通路为靶点的抗肿瘤中药研究进展.中国老年学杂志,2018,38(1)∶239～241.
[20]朱小勤,陈珮,黄中迪,等.中药调控血液肿瘤细胞周期的研究进展.上海中医药杂志,2015,49(9)∶83～86.