异常黑胆质成熟剂对异常黑胆质型肝癌病证大鼠模型候选基因表达的影响
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摘要
目的探讨异常黑胆质成熟剂(abnormal savda munziq,ASM)对异常黑胆质型肝癌病证大鼠模型候选基因表达水平的影响.方法选取♂清洁Wistar大鼠随机分为6个组.联合维吾尔医学理论及二乙基亚硝胺(diethylnitrosamine,DEN)诱导建立异常黑胆质型肝癌病证大鼠模型,并用ASM高(6.0 g/kg),中(3.0 g/kg),低(1.5 g/kg)不同剂量对模型组全程干预20 wk,观察肝脏组织形态学变化,并通过RT-qPCR验证表达谱芯片筛选出的部分差异表达候选基因.结果观察肝脏组织形态学变化发现,异常黑胆质型肝癌病证模型组成癌率明显高于对照肝癌组,ASM干预组成癌率明显低于异常黑胆质型肝癌病证模型组;芯片结果显示,与对照肝癌组比较,异常黑胆质型肝癌病证模型组上调表达基因438种、表达下调基因451种,对从异常黑胆质型肝癌病证模型组与ASM干预组表达变化相反的11种基因中选出6种进行RT-qPCR验证,结果发现,在大部分组间mRNA表达水平有差异并具有统计学意义(P<0.01,P<0.05).结论STAT3、CyclinD1、EGLN3、EMP1、NEFL、IGFALS基因可能是ASM抗癌的作用靶点.
AIM To identify the differentially expressed genes in an abnormal savda rat model of hepatocarcinoma treated with abnormal savda munziq(ASM).METHODS Male Wistar rats were randomly divided into six groups. An abnormal savda rat model of hepatocarcinoma was established with diethylnitrosamine(DEN) according to Uyghur medicine theory. Different doses of ASM(6.0, 3.0, and 1.5 g/kg) were used to intervene the model rats for 20 wk. The histological changes in hepatocarcinoma tissue were observed. The genes with differential expression were screened, several of which were investigated by RT-qPCR.RESULTS The histology of liver tissue showed that the cancerogenic rate was significantly higher in rats with abnormal savda than in those without. Cancerogenic rates in the ASM intervention groups were significantly lower than those in rats with abnormal savda. Compared with the control group, 438 genes were up-regulated and 451 down-regulated in abnormal savda rats with hepatocarcinoma. Among these differentially expressed genes, 11 showed converse expression patterns between the hepatocarcinoma group and ASM intervention groups. RT-qPCR verified that STAT3, CyclinD1, EGLN3, EMP1, NEFL, and IGFALS were differentially expressed in different groups(P < 0.01 or P < 0.05).CONCLUSION STAT3, CyclinD1, EGLN3, EMP1, NEFL, and IGFALS may be involved in the anti-cancer effects of ASM.
AIM To identify the differentially expressed genes in an abnormal savda rat model of hepatocarcinoma treated with abnormal savda munziq(ASM).METHODS Male Wistar rats were randomly divided into six groups. An abnormal savda rat model of hepatocarcinoma was established with diethylnitrosamine(DEN) according to Uyghur medicine theory. Different doses of ASM(6.0, 3.0, and 1.5 g/kg) were used to intervene the model rats for 20 wk. The histological changes in hepatocarcinoma tissue were observed. The genes with differential expression were screened, several of which were investigated by RT-qPCR.RESULTS The histology of liver tissue showed that the cancerogenic rate was significantly higher in rats with abnormal savda than in those without. Cancerogenic rates in the ASM intervention groups were significantly lower than those in rats with abnormal savda. Compared with the control group, 438 genes were up-regulated and 451 down-regulated in abnormal savda rats with hepatocarcinoma. Among these differentially expressed genes, 11 showed converse expression patterns between the hepatocarcinoma group and ASM intervention groups. RT-qPCR verified that STAT3, CyclinD1, EGLN3, EMP1, NEFL, and IGFALS were differentially expressed in different groups(P < 0.01 or P < 0.05).CONCLUSION STAT3, CyclinD1, EGLN3, EMP1, NEFL, and IGFALS may be involved in the anti-cancer effects of ASM.
引文
1 Chen W,Zheng R,Baade PD,Zhang S,Zeng H,Bray F,Jemal A,Yu XQ,He J.Cancer statistics in China,2015.CA Cancer J Clin 2016;66:115-132[PMID:26808342 DOI:10.3322/caac.21338]
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3 阻力皮卡尔·阿卜杜热合曼,哈木拉提·吾甫尔,斯坎德尔·白克力,那孜拉木·玉苏甫江,古丽尼格尔·雪合拉提,王延蛟.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.新疆医科大学学报2017;4:540-544
4 王延蛟,热沙来提·阿不都瓦衣特,哈木拉提·吾甫尔,热斯拉特·艾力木,阿尤甫江·阿布都热依木,斯坎德尔·白克力.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.科技导报2015;33:84-89
5 哈木拉提·吾甫尔,阿依努尔·买提斯迪克,努尔买买提·艾买提,阿迪力·阿不力米提.异常黑胆质证载体动物模型的建立及其自然恢复反证.新疆医科大学学报2006;29:910-914
6 哈木拉提·吾甫尔.维吾尔医气质,体液论及其现代研究.乌鲁木齐:新疆科学技术出版社,2003:34-37,131-133
7 张莉,哈木拉提·吾甫尔,玛依努尔·艾力,阿依努尔·麦提斯迪克,张建清.恶性肿瘤的维医分型及其神经内分泌免疫网络紊乱研究.中国中医基础医学杂志2008;14:119-121
8 阿依努尔·热合曼,艾尼瓦尔·托合提,伊利亚尔·木太里甫.维吾尔药异常黑胆质成熟剂和清除剂研究进展.科技视野2014;32:30-32
9 王延蛟,哈木拉提·吾甫尔,依马木·买买提依明,热斯拉特·艾力木,阿尤甫江·阿布都热依木,斯坎德尔·白克力.异常黑胆质型肝癌病证模型肝脏形态学研究.科技导报2014;32:74-78
10 Niu G,Bowman T,Huang M,Shivers S,Reintgen D,Daud A,Chang A,Kraker A,Jove R,Yu H.Roles of activated Src and Stat3 signaling in melanoma tumor cell growth.Oncogene 2002;21:7001-7010[PMID:12370822 DOI:10.1038/sj.onc.1205859]
11 苟泽鹏,王德盛,周亮,张勇,祝普利,卢鹏,孙伟.STAT3蛋白在肝癌组织中的表达及临床意义.临床肝胆病杂志2013;29:520-524
12 Lirdprapamongkol K,Sakurai H,Abdelhamed S,Yokoyama S,Athikomkulchai S,Viriyaroj A,Awale S,Ruchirawat S,Svasti J,Saiki I.Chrysin overcomes TRAIL resistance of cancer cells through Mcl-1 downregulation by inhibiting STAT3 phosphorylation.Int J Oncol 2013;43:329-337[PMID:23636231 DOI:10.3892/ijo.2013.1926]
13 Lee H,Pal SK,Reckamp K,Figlin RA,Yu H.STAT3:a target to enhance antitumor immune response.Curr Top Microbiol Immunol 2011;344:41-59[PMID:20517723 DOI:10.1007/82_2010_51]
14 张砚,陈卫刚,郑勇.CyclinD1及相关基因在食管癌中的研究进展.中华全科医学2011;9:443-444
15 Luo KJ,Hu Y,Wen J,Fu JH.CyclinD1,p53,E-cadherin,and VEGF discordant expression in paired regional metastatic lymph nodes of esophageal squamous cell carcinoma:a tissue array analysis.J Surg Oncol 2011;104:236-243[PMID:21480261 DOI:10.1002/jso.21921]
16 刘馨莲,李淑蓉,孙静,殷舞.细胞周期蛋白在食管癌中的表达及意义.中国实验诊断学2012;16:1249-1252
17 谷化平,黄勇,尚培中.细胞周期素D1和P16蛋白表达与胆管癌转移和预后的关系.肝胆胰外科杂志2013;25:219-231
18 Su C,Huang K,Sun L,Yang D,Zheng H,Gao C,Tong J,Zhang Q.Overexpression of the HIF hydroxylase PHD3 is a favorable prognosticator for gastric cancer.Med Oncol 2012;29:2710-2715[PMID:22290580 DOI:10.1007/s12032-012-0171-6]
19 Andersen S,Donnem T,Stenvold H,Al-Saad S,Al-Shibli K,Busund LT,Bremnes RM.Overexpression of the HIF hydroxylases PHD1,PHD2,PHD3 and FIH are individually and collectively unfavorable prognosticators for NSCLC survival.PLoS One 2011;6:e23847[PMID:21887331 DOI:10.1371/journal.pone.0023847]
20 Chen S,Zhang J,Li X,Luo X,Fang J,Chen H.The expression of prolyl hydroxylase domain enzymes are up-regulated and negatively correlated with Bcl-2 in non-small cell lung cancer.Mol Cell Biochem 2011;358:257-263[PMID:21748337 DOI:10.1007/s11010-011-0976-1]
21 Peurala E,Koivunen P,Bloigu R,Haapasaari KM,Jukkola-Vuorinen A.Expressions of individual PHDs associate with good prognostic factors and increased proliferation in breast cancer patients.Breast Cancer Res Treat 2012;133:179-188[PMID:21877141 DOI:10.1007/s10549-011-1750-5]
22 Sun GG,Wang YD,Cui DW,Cheng YJ,Hu WN.Epithelial membrane protein 1 negatively regulates cell growth and metastasis in colorectal carcinoma.World J Gastroenterol 2014;20:4001-4010[PMID:24744589 DOI:10.3748/wjg.v20.i14.4001]
23 Sun G,Zhao G,Lu Y,Wang Y,Yang C.Association of EMP1 with gastric carcinoma invasion,survival and prognosis.Int J Oncol 2014;45:1091-1098[PMID:24920167 DOI:10.3892/ijo.2014.2488]
24 Sun GG,Wang YD,Lu YF,Hu WN.EMP1,a member of a new family of antiproliferative genes in breast carcinoma.Tumour Biol 2014;35:3347-3354[PMID:24402572 DOI:10.1007/s13277-013-1441-4]
25 Sun GG,Wang YD,Cui DW,Cheng YJ,Hu WN.EMP1 regulates caspase-9 and VEGFC expression and suppresses prostate cancer cell proliferation and invasion.Tumour Biol 2014;35:3455-3462[PMID:24338711 DOI:10.1007/s13277-013-1456-x]
26 Imbert A,Chaffanet M,Essioux L,Noguchi T,Ad閘a飀e J,Kerangueven F,Le Paslier D,Bona飔i-PelliéC,Sobol H,Birnbaum D,Pébusque MJ.Integrated map of the chromosome 8p12-p21 region,a region involved in human cancers and Werner syndrome.Genomics 1996;32:29-38[PMID:8786118 DOI:10.1006/geno.1996.0073]
27 Li XQ,Li L,Xiao CH,Feng YM.NEFL mRNA expression level is a prognostic factor for early-stage breast cancer patients.PLoS One 2012;7:e31146[PMID:22319610 DOI:10.1371/journal.pone.0031146]
28 Kang S,Kim B,Park SB,Jeong G,Kang HS,Liu R,Kim SJ.Stage-specific methylome screen identifies that NEFL is downregulated by promoter hypermethylation in breast cancer.Int J Oncol 2013;43:1659-1665[PMID:24026393 DOI:10.3892/ijo.2013.2094]
29 Peng G,Yuan X,Yuan J,Liu Q,Dai M,Shen C,Ma J,Liao Y,Jiang W.miR-25 promotes glioblastoma cell proliferation and invasion by directly targeting NEFL.Mol Cell Biochem 2015;409:103-111[PMID:26209061 DOI:10.1007/s11010-015-2516-x]
30 鲍丽颖,黄耕培.GH-IGF-I轴与骨发育的研究进展.天津体育学院学报2002;17:60-62
31 郑凯迪.斑马鱼Igfals的表达、转录调控及功能研究.重庆:西南大学,2010:12-14
32 Grandone A,Miraglia del Giudice E,Cirillo G,Abbondanza C,Cioffi M,Romano T,Micillo F,Marzuillo P,Perrone L.Clinical features of a new acid-labile subunit(IGFALS)heterozygous mutation:anthropometric and biochemical characterization and response to growth hormone administration.Horm Res Paediatr 2014;81:67-72[PMID:24356109 DOI:10.1159/000355017]
2 胡汉华,盛磊,哈木拉提·吾甫尔,刘翠,熊文娟,吴彩云,姑丽尼格尔·库来西.维药异常黑胆质成熟剂的抗肿瘤作用及其对细胞迁移的影响.科技导报2011;29:62-65
3 阻力皮卡尔·阿卜杜热合曼,哈木拉提·吾甫尔,斯坎德尔·白克力,那孜拉木·玉苏甫江,古丽尼格尔·雪合拉提,王延蛟.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.新疆医科大学学报2017;4:540-544
4 王延蛟,热沙来提·阿不都瓦衣特,哈木拉提·吾甫尔,热斯拉特·艾力木,阿尤甫江·阿布都热依木,斯坎德尔·白克力.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.科技导报2015;33:84-89
5 哈木拉提·吾甫尔,阿依努尔·买提斯迪克,努尔买买提·艾买提,阿迪力·阿不力米提.异常黑胆质证载体动物模型的建立及其自然恢复反证.新疆医科大学学报2006;29:910-914
6 哈木拉提·吾甫尔.维吾尔医气质,体液论及其现代研究.乌鲁木齐:新疆科学技术出版社,2003:34-37,131-133
7 张莉,哈木拉提·吾甫尔,玛依努尔·艾力,阿依努尔·麦提斯迪克,张建清.恶性肿瘤的维医分型及其神经内分泌免疫网络紊乱研究.中国中医基础医学杂志2008;14:119-121
8 阿依努尔·热合曼,艾尼瓦尔·托合提,伊利亚尔·木太里甫.维吾尔药异常黑胆质成熟剂和清除剂研究进展.科技视野2014;32:30-32
9 王延蛟,哈木拉提·吾甫尔,依马木·买买提依明,热斯拉特·艾力木,阿尤甫江·阿布都热依木,斯坎德尔·白克力.异常黑胆质型肝癌病证模型肝脏形态学研究.科技导报2014;32:74-78
10 Niu G,Bowman T,Huang M,Shivers S,Reintgen D,Daud A,Chang A,Kraker A,Jove R,Yu H.Roles of activated Src and Stat3 signaling in melanoma tumor cell growth.Oncogene 2002;21:7001-7010[PMID:12370822 DOI:10.1038/sj.onc.1205859]
11 苟泽鹏,王德盛,周亮,张勇,祝普利,卢鹏,孙伟.STAT3蛋白在肝癌组织中的表达及临床意义.临床肝胆病杂志2013;29:520-524
12 Lirdprapamongkol K,Sakurai H,Abdelhamed S,Yokoyama S,Athikomkulchai S,Viriyaroj A,Awale S,Ruchirawat S,Svasti J,Saiki I.Chrysin overcomes TRAIL resistance of cancer cells through Mcl-1 downregulation by inhibiting STAT3 phosphorylation.Int J Oncol 2013;43:329-337[PMID:23636231 DOI:10.3892/ijo.2013.1926]
13 Lee H,Pal SK,Reckamp K,Figlin RA,Yu H.STAT3:a target to enhance antitumor immune response.Curr Top Microbiol Immunol 2011;344:41-59[PMID:20517723 DOI:10.1007/82_2010_51]
14 张砚,陈卫刚,郑勇.CyclinD1及相关基因在食管癌中的研究进展.中华全科医学2011;9:443-444
15 Luo KJ,Hu Y,Wen J,Fu JH.CyclinD1,p53,E-cadherin,and VEGF discordant expression in paired regional metastatic lymph nodes of esophageal squamous cell carcinoma:a tissue array analysis.J Surg Oncol 2011;104:236-243[PMID:21480261 DOI:10.1002/jso.21921]
16 刘馨莲,李淑蓉,孙静,殷舞.细胞周期蛋白在食管癌中的表达及意义.中国实验诊断学2012;16:1249-1252
17 谷化平,黄勇,尚培中.细胞周期素D1和P16蛋白表达与胆管癌转移和预后的关系.肝胆胰外科杂志2013;25:219-231
18 Su C,Huang K,Sun L,Yang D,Zheng H,Gao C,Tong J,Zhang Q.Overexpression of the HIF hydroxylase PHD3 is a favorable prognosticator for gastric cancer.Med Oncol 2012;29:2710-2715[PMID:22290580 DOI:10.1007/s12032-012-0171-6]
19 Andersen S,Donnem T,Stenvold H,Al-Saad S,Al-Shibli K,Busund LT,Bremnes RM.Overexpression of the HIF hydroxylases PHD1,PHD2,PHD3 and FIH are individually and collectively unfavorable prognosticators for NSCLC survival.PLoS One 2011;6:e23847[PMID:21887331 DOI:10.1371/journal.pone.0023847]
20 Chen S,Zhang J,Li X,Luo X,Fang J,Chen H.The expression of prolyl hydroxylase domain enzymes are up-regulated and negatively correlated with Bcl-2 in non-small cell lung cancer.Mol Cell Biochem 2011;358:257-263[PMID:21748337 DOI:10.1007/s11010-011-0976-1]
21 Peurala E,Koivunen P,Bloigu R,Haapasaari KM,Jukkola-Vuorinen A.Expressions of individual PHDs associate with good prognostic factors and increased proliferation in breast cancer patients.Breast Cancer Res Treat 2012;133:179-188[PMID:21877141 DOI:10.1007/s10549-011-1750-5]
22 Sun GG,Wang YD,Cui DW,Cheng YJ,Hu WN.Epithelial membrane protein 1 negatively regulates cell growth and metastasis in colorectal carcinoma.World J Gastroenterol 2014;20:4001-4010[PMID:24744589 DOI:10.3748/wjg.v20.i14.4001]
23 Sun G,Zhao G,Lu Y,Wang Y,Yang C.Association of EMP1 with gastric carcinoma invasion,survival and prognosis.Int J Oncol 2014;45:1091-1098[PMID:24920167 DOI:10.3892/ijo.2014.2488]
24 Sun GG,Wang YD,Lu YF,Hu WN.EMP1,a member of a new family of antiproliferative genes in breast carcinoma.Tumour Biol 2014;35:3347-3354[PMID:24402572 DOI:10.1007/s13277-013-1441-4]
25 Sun GG,Wang YD,Cui DW,Cheng YJ,Hu WN.EMP1 regulates caspase-9 and VEGFC expression and suppresses prostate cancer cell proliferation and invasion.Tumour Biol 2014;35:3455-3462[PMID:24338711 DOI:10.1007/s13277-013-1456-x]
26 Imbert A,Chaffanet M,Essioux L,Noguchi T,Ad閘a飀e J,Kerangueven F,Le Paslier D,Bona飔i-PelliéC,Sobol H,Birnbaum D,Pébusque MJ.Integrated map of the chromosome 8p12-p21 region,a region involved in human cancers and Werner syndrome.Genomics 1996;32:29-38[PMID:8786118 DOI:10.1006/geno.1996.0073]
27 Li XQ,Li L,Xiao CH,Feng YM.NEFL mRNA expression level is a prognostic factor for early-stage breast cancer patients.PLoS One 2012;7:e31146[PMID:22319610 DOI:10.1371/journal.pone.0031146]
28 Kang S,Kim B,Park SB,Jeong G,Kang HS,Liu R,Kim SJ.Stage-specific methylome screen identifies that NEFL is downregulated by promoter hypermethylation in breast cancer.Int J Oncol 2013;43:1659-1665[PMID:24026393 DOI:10.3892/ijo.2013.2094]
29 Peng G,Yuan X,Yuan J,Liu Q,Dai M,Shen C,Ma J,Liao Y,Jiang W.miR-25 promotes glioblastoma cell proliferation and invasion by directly targeting NEFL.Mol Cell Biochem 2015;409:103-111[PMID:26209061 DOI:10.1007/s11010-015-2516-x]
30 鲍丽颖,黄耕培.GH-IGF-I轴与骨发育的研究进展.天津体育学院学报2002;17:60-62
31 郑凯迪.斑马鱼Igfals的表达、转录调控及功能研究.重庆:西南大学,2010:12-14
32 Grandone A,Miraglia del Giudice E,Cirillo G,Abbondanza C,Cioffi M,Romano T,Micillo F,Marzuillo P,Perrone L.Clinical features of a new acid-labile subunit(IGFALS)heterozygous mutation:anthropometric and biochemical characterization and response to growth hormone administration.Horm Res Paediatr 2014;81:67-72[PMID:24356109 DOI:10.1159/000355017]