异常黑胆质成熟剂对异常黑胆质型肝癌病证模型大鼠Id4和p21 mRNA表达水平的影响
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摘要
目的探讨维药异常黑胆质成熟剂在异常黑胆质型肝癌病证大鼠模型中的抗肝癌作用。方法 90只清洁级雄性Wistar大鼠按随机数字表法分成正常组、对照肝癌组、异黑肝癌组和成熟剂高、中、低剂量组6组,每组各15只。正常组正常饲养,对照肝癌组正常饲养21 d后给予二乙基亚硝胺(DEN)20周诱导肝癌。异黑肝癌组和成熟剂高、中、低剂量组大鼠通过干寒属性饲料、干寒气候环境、足底电刺激,夹尾以及强迫游泳等多因素复合作用21 d建立异常黑胆质证载体动物模型,再加用DEN20周诱导建立异常黑胆质型肝癌病证模型;DEN诱导的同时,成熟剂高、中、低剂量组用异常黑胆质成熟剂按高、中、低(6.0、3.0、1.5 g/kg)剂量进行干预。提取肝组织中m RNA,利用基因芯片技术筛选出的大鼠肝脏差异表达的Id4和p21候选基因并用RT-qPCR法验证。结果与正常组比较,对照肝癌组肝组织Id4基因表达下调(P<0.01);与对照肝癌组比较,异黑肝癌组肝组织Id4基因表达下调(P<0.01);与异黑肝癌组相比,成熟剂高、中、低剂量肝组织Id4基因表达均上调,其中成熟剂高、中剂量组升高显著(P<0.01)。与正常组相比,对照肝癌组肝组织p21基因表达上调(P<0.01);与对照肝癌组比较,异黑肝癌组肝组织p21基因表达上调(P<0.05);与异黑肝癌组相比,成熟剂高、中、低剂量肝组织p21基因表达均下调,其中成熟剂中、低剂量组下降明显(P<0.01)。结论异常黑胆质成熟剂可影响异常黑胆质型肝癌病证模型大鼠肝脏Id4和p21基因的m RNA表达水平。
Objective To investigate the anticancer role of Uyghur medicine Abnormal Savda Munziq(ASM) in abnormal savda hepatocellular carcinoma rat models. Methods 90 male Wistar rats were divided into 6 groups, including normal group, control hepatic carcinoma group, abnormal savda hepatocellular carcinoma group and ASM high, middle and low dosage group by random number table, with 15 cases in each group. The normal group rats were placed in normative feeding environment. After 21 days of feeding in laboratory environment, the control hepatic carcinoma group rats were induced by diethylnitrosamine(DEN) 20 weeks to establish the liver cancer rat model. The rats in the abnormal savda hepatocellular carcinoma group, ASM high, middle and low dosage group were established abnormal black bile models, through dry cold property feed, dry cold climate environment, plantar electrical stimulation, tail and forced swimming, etc; after 21 days they were induced by DEN for 20 weeks to establish the abnormal savda hepatocellular carcinoma models. At the same time, the rats in the ASM high, middle and low dosage group were intervened by 6.0,3.0, 1.5 g/kg of ASM. The m RNA was extracted from liver tissue and Id4, p21 gene were verified by RT-qPCR after screened by gene chip technology. Results The Id4 gene expression in liver tissue was down-regulated in control hep-atic carcinoma group,compared with the normal control group(P < 0.01). The Id4 gene expression in liver tissue was down-regulated in abnormal savda syndromeliver cancer group, compared with the control hepatic carcinoma group(P < 0.01). Compared with the abnormal savda hepatocellular carcinoma grou p, the expression of Id4 gene in liver tissue was up-regulated all inASM high, middle and low dosage group, the high and middle dosage groups were obviously up-regulated(P < 0.01).Compared with the normal group, the liver tissue p21 gene expression was up-regulated in control hepatic carcinoma group(P < 0.01). The p21 gene expression in liver tissue wasup-regulated in abnormal savda syndromeliver cancer group, compared with the control hepatic carcinoma group(P < 0.05). Compared with the abnormal savda hepatocellular carcinoma group, the expressions of p21 gene were down-regulated all in ASM high, middle and low dosage group, the middle and low dosages group were dramatic decline(P < 0.01). Conclusion The ASM could have an impact on m RNA expressions of Id4 and p21 gene in abnormal savda hepatocellular carcinoma rat models.
Objective To investigate the anticancer role of Uyghur medicine Abnormal Savda Munziq(ASM) in abnormal savda hepatocellular carcinoma rat models. Methods 90 male Wistar rats were divided into 6 groups, including normal group, control hepatic carcinoma group, abnormal savda hepatocellular carcinoma group and ASM high, middle and low dosage group by random number table, with 15 cases in each group. The normal group rats were placed in normative feeding environment. After 21 days of feeding in laboratory environment, the control hepatic carcinoma group rats were induced by diethylnitrosamine(DEN) 20 weeks to establish the liver cancer rat model. The rats in the abnormal savda hepatocellular carcinoma group, ASM high, middle and low dosage group were established abnormal black bile models, through dry cold property feed, dry cold climate environment, plantar electrical stimulation, tail and forced swimming, etc; after 21 days they were induced by DEN for 20 weeks to establish the abnormal savda hepatocellular carcinoma models. At the same time, the rats in the ASM high, middle and low dosage group were intervened by 6.0,3.0, 1.5 g/kg of ASM. The m RNA was extracted from liver tissue and Id4, p21 gene were verified by RT-qPCR after screened by gene chip technology. Results The Id4 gene expression in liver tissue was down-regulated in control hep-atic carcinoma group,compared with the normal control group(P < 0.01). The Id4 gene expression in liver tissue was down-regulated in abnormal savda syndromeliver cancer group, compared with the control hepatic carcinoma group(P < 0.01). Compared with the abnormal savda hepatocellular carcinoma grou p, the expression of Id4 gene in liver tissue was up-regulated all inASM high, middle and low dosage group, the high and middle dosage groups were obviously up-regulated(P < 0.01).Compared with the normal group, the liver tissue p21 gene expression was up-regulated in control hepatic carcinoma group(P < 0.01). The p21 gene expression in liver tissue wasup-regulated in abnormal savda syndromeliver cancer group, compared with the control hepatic carcinoma group(P < 0.05). Compared with the abnormal savda hepatocellular carcinoma group, the expressions of p21 gene were down-regulated all in ASM high, middle and low dosage group, the middle and low dosages group were dramatic decline(P < 0.01). Conclusion The ASM could have an impact on m RNA expressions of Id4 and p21 gene in abnormal savda hepatocellular carcinoma rat models.
引文
[1]陈启亮,唐东昕,龙奉玺.中医药对肝癌治疗的研究进展[J].贵阳中医学院学报,2016,38(2):99-101.
[2]刘莹莹,邓皖利,刘文先,等.异常黑胆质成熟剂治疗肿瘤的研究进展[J].中华中医药杂志,2015,30(1):156-158.
[3]阿尤甫江·阿布都热依木,斯坎德尔·白克力,王延娇,等.HPAA组织结构和功能的变化在异常黑胆质型肝癌病证模型中的作用研究[J].新疆医科大学学报,2015,38(3):273-277.
[4]王延蛟,哈木拉提·吾甫尔,依马木·买买提依明,等.异常黑胆质型肝癌病证模型肝脏形态学研究[J].科技导报,2014,32(14):74-78.
[5]哈木拉提·吾甫尔.能够使异常黑胆质型体液成熟和清除的药物及其制备方法:中国,02130082.8[P].2003-04-06.
[6]Pankaj S,Swathi C,Jaideep C.Inhibitor of differentiation 4(Id4)acts as an inhibitor of ID-1,-2 and-3 and promotes basic Helix Loop Helix(b HLH)E47 DNA binding and transcriptional Activity[J].Biochimie,2015,112:139-150.
[7]Stefania D,Federica G,Sabrina S,et al.ID4:a new player in the cancer arena[J].Oncotarget,2010,1(1):48-58.
[8]Derrick JM,Divya P,Jugal J,et al.ID4 regulates transcriptional activity of wild type and mutant p53 via K373acetylation[J].Oncotarget,2017,8(2):2536-2549.
[9]Jason PC,Ashley EK,Swathi C,et al.Id4 promotes senescence and sensitivity to doxorubicin-induced apoptosis in DU145 prostate cancer cells[J].Anticancer Res,2013,33(10):4271-4278.
[10]Rajiv RM,Brandie RM,Govindaraj A,et al.Characterization of inhibitor of differentiation(Id)proteins in human cornea[J].Exp Eye Res,2016,146:145-153.
[11]Divya P,Derrick JM,Jason C,et al.Inhibitor of differentiation 4(id4):from development to cancer[J].Biochim Biophys Acta,2015,1855(1):92-103
[12]Gao XZ,Zhao WG,Wang GN,et al.Inhibitor of DNA binding 4 functions as a tumor suppressor and is targetable by 5-aza-2′-deoxycytosine with potential therapeutic significance in Burkitt′s lymphoma[J].Mol Med Rep,2016,13(2):1269-1274.
[13]Kang H,Wang X,Gao L,et al.Clinical implications of the quantitative detection of ID4 gene methylation in myelodysplastic syndrome[J].Eur J Med Res,2015,20:16.
[14]Zhang Y,Zhan g B,Fang J,et al.Hypomethylation of DNA-binding inhibitor 4 serves as a potential biomarker in distinguishing acquired tamoxifen-refractory breast cancer[J].Int J Clin Exp Pathol,2015,8(8):9500-9505.
[15]Vinarskaja A,Goering W,Ingenwerth M,et al.ID4 is frequently downregulated and partially hypermethylated in prostate cancer[J].World J Urol,2012,30(3):319-325.
[16]Qi K,Li Y,Li X,et al.Id4 promotes cisplatin resistance in lung cancer through the p38 MAPK pathway[J].Anti Cancer Drugs,2016,27(10):970-978.
[17]Zhang Y,Zhang LX,Liu XQ,et al.Id4 promotes cell proliferation in hepatocellular carcinoma[J].Chin J Cancer,2017,36(1):19.
[18]Karimian A,Ahmadi Y,Yousefi B.Multiple functions of p21 in cell cycle,apoptosis and transcriptional regulation after DNA damage[J].DNA Repair,2016,42:63-71.
[19]Dolezalova D,Mraz M,Barta T,et al.Micro RNAs regulate p21(Waf1/Cip1)protein expression and the DNA dam age response in human embryonic stem cells[J].Stem Cells,2012,30(7):1362-1372.
[20]Uryga A,Gray K,Bennett M.DNA damage and repair in vascular disease[J].Annu Rev Physiol,2016,78:45-66.
[21]许杨,曾小云.P21基因表达与肝细胞肝癌关系的病例对照研究[J].广西医科大学学报,2012,29(1):7-9.
[22]祖力皮喀尔·阿卜杜热合曼,哈木拉提·吾甫尔,斯坎德尔·白克力,等.异常黑胆质成熟剂对多因素诱发肝癌大鼠模型中p53和p21基因表达的影响[J].新疆医科大学学报,2017,40(4):540-544.
[23]买买提·努尔艾合提,厉蓓,董竞成.维吾尔族传统医学异常黑胆质证的研究概况[J].中国民族医药杂志,2013,19(12):44-49.
[24]王延蛟,热沙来提·阿卜都瓦衣特,哈木拉提·吾甫尔,等.异常黑胆质成熟剂对异常黑胆质型肝癌病证大鼠模型肝脏形态学的影响[J].科技导报,2015,33(11):84-89.
[2]刘莹莹,邓皖利,刘文先,等.异常黑胆质成熟剂治疗肿瘤的研究进展[J].中华中医药杂志,2015,30(1):156-158.
[3]阿尤甫江·阿布都热依木,斯坎德尔·白克力,王延娇,等.HPAA组织结构和功能的变化在异常黑胆质型肝癌病证模型中的作用研究[J].新疆医科大学学报,2015,38(3):273-277.
[4]王延蛟,哈木拉提·吾甫尔,依马木·买买提依明,等.异常黑胆质型肝癌病证模型肝脏形态学研究[J].科技导报,2014,32(14):74-78.
[5]哈木拉提·吾甫尔.能够使异常黑胆质型体液成熟和清除的药物及其制备方法:中国,02130082.8[P].2003-04-06.
[6]Pankaj S,Swathi C,Jaideep C.Inhibitor of differentiation 4(Id4)acts as an inhibitor of ID-1,-2 and-3 and promotes basic Helix Loop Helix(b HLH)E47 DNA binding and transcriptional Activity[J].Biochimie,2015,112:139-150.
[7]Stefania D,Federica G,Sabrina S,et al.ID4:a new player in the cancer arena[J].Oncotarget,2010,1(1):48-58.
[8]Derrick JM,Divya P,Jugal J,et al.ID4 regulates transcriptional activity of wild type and mutant p53 via K373acetylation[J].Oncotarget,2017,8(2):2536-2549.
[9]Jason PC,Ashley EK,Swathi C,et al.Id4 promotes senescence and sensitivity to doxorubicin-induced apoptosis in DU145 prostate cancer cells[J].Anticancer Res,2013,33(10):4271-4278.
[10]Rajiv RM,Brandie RM,Govindaraj A,et al.Characterization of inhibitor of differentiation(Id)proteins in human cornea[J].Exp Eye Res,2016,146:145-153.
[11]Divya P,Derrick JM,Jason C,et al.Inhibitor of differentiation 4(id4):from development to cancer[J].Biochim Biophys Acta,2015,1855(1):92-103
[12]Gao XZ,Zhao WG,Wang GN,et al.Inhibitor of DNA binding 4 functions as a tumor suppressor and is targetable by 5-aza-2′-deoxycytosine with potential therapeutic significance in Burkitt′s lymphoma[J].Mol Med Rep,2016,13(2):1269-1274.
[13]Kang H,Wang X,Gao L,et al.Clinical implications of the quantitative detection of ID4 gene methylation in myelodysplastic syndrome[J].Eur J Med Res,2015,20:16.
[14]Zhang Y,Zhan g B,Fang J,et al.Hypomethylation of DNA-binding inhibitor 4 serves as a potential biomarker in distinguishing acquired tamoxifen-refractory breast cancer[J].Int J Clin Exp Pathol,2015,8(8):9500-9505.
[15]Vinarskaja A,Goering W,Ingenwerth M,et al.ID4 is frequently downregulated and partially hypermethylated in prostate cancer[J].World J Urol,2012,30(3):319-325.
[16]Qi K,Li Y,Li X,et al.Id4 promotes cisplatin resistance in lung cancer through the p38 MAPK pathway[J].Anti Cancer Drugs,2016,27(10):970-978.
[17]Zhang Y,Zhang LX,Liu XQ,et al.Id4 promotes cell proliferation in hepatocellular carcinoma[J].Chin J Cancer,2017,36(1):19.
[18]Karimian A,Ahmadi Y,Yousefi B.Multiple functions of p21 in cell cycle,apoptosis and transcriptional regulation after DNA damage[J].DNA Repair,2016,42:63-71.
[19]Dolezalova D,Mraz M,Barta T,et al.Micro RNAs regulate p21(Waf1/Cip1)protein expression and the DNA dam age response in human embryonic stem cells[J].Stem Cells,2012,30(7):1362-1372.
[20]Uryga A,Gray K,Bennett M.DNA damage and repair in vascular disease[J].Annu Rev Physiol,2016,78:45-66.
[21]许杨,曾小云.P21基因表达与肝细胞肝癌关系的病例对照研究[J].广西医科大学学报,2012,29(1):7-9.
[22]祖力皮喀尔·阿卜杜热合曼,哈木拉提·吾甫尔,斯坎德尔·白克力,等.异常黑胆质成熟剂对多因素诱发肝癌大鼠模型中p53和p21基因表达的影响[J].新疆医科大学学报,2017,40(4):540-544.
[23]买买提·努尔艾合提,厉蓓,董竞成.维吾尔族传统医学异常黑胆质证的研究概况[J].中国民族医药杂志,2013,19(12):44-49.
[24]王延蛟,热沙来提·阿卜都瓦衣特,哈木拉提·吾甫尔,等.异常黑胆质成熟剂对异常黑胆质型肝癌病证大鼠模型肝脏形态学的影响[J].科技导报,2015,33(11):84-89.