T0901317通过法尼醇X受体调节人肠癌细胞载脂蛋白M表达

载脂蛋白M(ApoM)是一种脂质运载蛋白家族的血浆蛋白质,主要存在于HDL中[1].本研究旨在观察TO901317通过法尼醇X受体调节人肠癌细胞载脂蛋白M表达.     

人锌指蛋白23在原发性肝细胞肝癌中的表达及临床意义

Objective To detect the expression level of human zinc finger 23 (ZNF23) in hepatocellular carcinoma tissue samples and HepG2 cell lines and investigate the relationship between hZNF23 expression and clinicopathological characteristics of HCC and cell apoptosis. Methods The expression levels of hZNF23 and GAPDH mRNA in 37 cases of HCC were measured by real-time RT-PCR. The association between the expression of hZNF23 and the clinicopathological characteristics of HCC was analyzed. Cultured HepG2 cells were divided into 4 groups ( control group, 1.25 μg/ml , 2.5 μg/ml and 5 μg/ml cisplatin)or 6 groups( control group, 1.25 μg/ml, 2.5 μg/ml, 5 μg/ml, 10 μg/ml and 20 μg/ml cisplatin). MTT method was employed to evaluate cell proliferation. Annexin V-FITC assay was used to assess percentage of apoptotic HepG2 cells. The expression levels of hZNF23 and GAPDH mRNA of HepG2 cells after apoptosis induced by cisplatin with a series of concentrations were measured by real-time RT-PCR.Results The median ( quartile1, quartile 3) expression levels of hZNF23 mRNA in 37 HCC tissue samples and adjacent tissue samples were 8.84 (3.59-15.05), 22.20 ( 13.85-42.90 ), respectively. There was significant difference ( U = 259.5, P ＜ 0.01 ). The median ( quartile1, quartile 3 ) expression levels of hZNF23 mRNA in cancer tissue samples with Edmondson stage Ⅰ + Ⅱ [12.80(4.80-19.50)] was much higher than those in stage Ⅲ + Ⅳ [5.01 ( 2.88-11.68 ), U = 99.00, P ＜ 0.05] The median ( quartile1,quartile 3 ) expression levels of hZNF23 mRNA in patients with and without hepatic cirrhosis were 9.92(3.80-15.25) , 3.21 (2.78-3.60), respectively. The median ( quartile1, quartile 3 ) expression levels of hZNF23 mRNA in HBV infection and non-infection patients was 9.09(3.72-15.25 ), 2.48 (1.79-12.10),respectively. There was no significant difference between groups with and without hepatic cirrhosis and between HBV infection and non-infection groups( U = 16. 00 and 24.00, P ＞0.05 ). MTT assay indicated that cisplatin significantly inhibited HepG2 cells proliferation in a dose-dependent manner. Annexin V-FITC/PI assay showed that HepG2 cells apoptosis rates were (0.9 ± 0.2 ) %, ( 4. 2 ± 0.3 ) %, ( 9.8 ± 4. 3 ) %,(23.0 ± 6.0)%, respectively. Cisplatin significantly induced HepG2 cells apoptosis in a dose-dependent manner( F = 27.89, P ＜ 0.01 ). The expression levels of hZNF23 mRNA in cisplatin groups [( 10.39 ±3.08) × 10-5, (24.10 ± 2.09) × 10-5, (6.90 ± 2.24) × 10-4] were significantly lower than that of the controlgroup[(94.80±1.80) ×10-5, F=6.027, P＜0.01]. Conclusions The expression level hZNF23 mRNA is related to Edmondson stage of HCC. The apoptosis effect of cisplatin on HepG2 cells may be associated with the upregulation of hZNF23.     

实时荧光定量聚合酶链反应检测人锌指蛋白23方法的构建

目的建立检测人锌指蛋白23(human zinc finger 23,ZNF23)的实时荧光定量聚合酶链反应(PCR)技术。方法采用Primer Express Versions 3.0设计引物及TaqMan探针,检测ZNF23基因。将扩增的ZNF23基因片段纯化后与pMD19-T载体连接,克隆构建含ZNF23基因片段的重组质粒,作为定量检测ZNF23基因表达的标准品。结果 PCR扩增产物经测序分析证实为ZNF23特异性片段。本法灵敏度为650 copies/μL,线性范围为(6.50×102)-(6.50×107)copies/μL,相关系数R2为0.999,扩增效率E为97.28%,批间变异系数为0.78%～7.53%。结论成功建立了检测人ZNF23的实时荧光定量PCR方法,且该方法特异性好、灵敏度高、通量高。     

颈动脉粥样硬化患者MT2A-838G/C和MCP-1-2518G/A基因多态性分析

目的: 探讨苏南地区汉族人群MT2A-838G/C和MCP-1-2518G/A基因多态性与颈动脉粥样硬化的相关性。方法： 采用碱基淬灭探针技术检测103例有颈动脉斑块患者和46例正常对照的MT2A基因-838位点和MCP-1基因-2518位点基因型分布及基因频率,并评价基因多态性与颈动脉粥样硬化易感性之间的相关性。结果： 在对照组和病例组中,MT2A 3种基因型GG、GC、CC的频率分别为52.17%、36.96%、10.87%和50.49%、36.89%、12.62%（P>0.05）,等位基因G、C的频率分别为70.65%、29.35%和68.93%、31.07%（P>0.05）;MCP-1 3种基因型AA、AG、GC在对照组和病例组中的频率分别为15.22%、52.17%、32.61%和20.39%、44.66%、34.95%（P>0.05）,等位基因A、G的频率分别为41.30%、58.70%和42.72%、57.28%（P>0.05）。结论： MT2A-838G/C及MCP-1-2518G/A基因多态性可能不是苏南汉族人群发生颈动脉粥样硬化的独立危险因素。     

ATP结合盒转运子A1调控载脂蛋白M表达的通路

目的研究人肝癌细胞株HepG2中ATP结合盒转运子A1（ABCA1）是否通过肝受体同系物（LRH1）调节载脂蛋白M（apoM）的表达。方法分别用肝X受体激动剂GW3965或ABCA1阻滞剂DIDS作用于HepG2,应用RT-PCR法检测ABCA1、LRH1、apoM及apoAI的mRNA水平。结果与DMSO组比较,GW3965能显著增加ABCA1mRNA的表达（P〈0.01）,且呈剂量依赖性下调LRH1（P〈0.01）,但apoM和apoAI mRNA水平无明显变化（P〉0.05）;与GW3965单独作用组比较,GW3965与DIDS联合用药组apoM与apoAI的基因表达均明显下调（P〈0.05）,但ABCA1和LRH1水平均无明显变化（P〉0.05）。结论 ABCA1不通过LRH1调节apoM的表达。     

人锌指蛋白23在原发性肝细胞肝癌中的表达及临床意义

Objective To detect the expression level of human zinc finger 23 (ZNF23) in hepatocellular carcinoma tissue samples and HepG2 cell lines and investigate the relationship between hZNF23 expression and clinicopathological characteristics of HCC and cell apoptosis. Methods The expression levels of hZNF23 and GAPDH mRNA in 37 cases of HCC were measured by real-time RT-PCR. The association between the expression of hZNF23 and the clinicopathological characteristics of HCC was analyzed. Cultured HepG2 cells were divided into 4 groups ( control group, 1.25 μg/ml , 2.5 μg/ml and 5 μg/ml cisplatin)or 6 groups( control group, 1.25 μg/ml, 2.5 μg/ml, 5 μg/ml, 10 μg/ml and 20 μg/ml cisplatin). MTT method was employed to evaluate cell proliferation. Annexin V-FITC assay was used to assess percentage of apoptotic HepG2 cells. The expression levels of hZNF23 and GAPDH mRNA of HepG2 cells after apoptosis induced by cisplatin with a series of concentrations were measured by real-time RT-PCR.Results The median ( quartile1, quartile 3) expression levels of hZNF23 mRNA in 37 HCC tissue samples and adjacent tissue samples were 8.84 (3.59-15.05), 22.20 ( 13.85-42.90 ), respectively. There was significant difference ( U = 259.5, P ＜ 0.01 ). The median ( quartile1, quartile 3 ) expression levels of hZNF23 mRNA in cancer tissue samples with Edmondson stage Ⅰ + Ⅱ [12.80(4.80-19.50)] was much higher than those in stage Ⅲ + Ⅳ [5.01 ( 2.88-11.68 ), U = 99.00, P ＜ 0.05] The median ( quartile1,quartile 3 ) expression levels of hZNF23 mRNA in patients with and without hepatic cirrhosis were 9.92(3.80-15.25) , 3.21 (2.78-3.60), respectively. The median ( quartile1, quartile 3 ) expression levels of hZNF23 mRNA in HBV infection and non-infection patients was 9.09(3.72-15.25 ), 2.48 (1.79-12.10),respectively. There was no significant difference between groups with and without hepatic cirrhosis and between HBV infection and non-infection groups( U = 16. 00 and 24.00, P ＞0.05 ). MTT assay indicated that cisplatin significantly inhibited HepG2 cells proliferation in a dose-dependent manner. Annexin V-FITC/PI assay showed that HepG2 cells apoptosis rates were (0.9 ± 0.2 ) %, ( 4. 2 ± 0.3 ) %, ( 9.8 ± 4. 3 ) %,(23.0 ± 6.0)%, respectively. Cisplatin significantly induced HepG2 cells apoptosis in a dose-dependent manner( F = 27.89, P ＜ 0.01 ). The expression levels of hZNF23 mRNA in cisplatin groups [( 10.39 ±3.08) × 10-5, (24.10 ± 2.09) × 10-5, (6.90 ± 2.24) × 10-4] were significantly lower than that of the controlgroup[(94.80±1.80) ×10-5, F=6.027, P＜0.01]. Conclusions The expression level hZNF23 mRNA is related to Edmondson stage of HCC. The apoptosis effect of cisplatin on HepG2 cells may be associated with the upregulation of hZNF23.     

载脂蛋白M在心肌细胞缺氧/复氧损伤中的表达差异及其作用

目的:探讨载脂蛋白M(apolipoprotein M,ApoM)在大鼠H9C2心肌细胞缺氧/复氧(hypoxia/reoxygenation,H/R)损伤中的表达差异及作用.方法:利用安宁包缺氧体系构建大鼠H9C2心肌细胞H/R损伤模型,采用CCK-8、细胞凋亡检测试剂盒、caspase-3活性检测试剂盒检测H9C2...     

环孢素A联合苯基棕榈酰胺吗啡丙醇对K562/AO2细胞GCS基因表达的影响及对逆转多药耐药

目的 研究环孢素A(CsA)和苯基棕榈酰胺吗啡丙醇(PPMP)联合应用对人白血病多药耐药细胞株K562/AO2的葡萄糖神经酰胺合酶(GCS)基因表达的影响及对白血病多药耐药的逆转作用,探索逆转白血病细胞耐药的策略.方法 应用Alamar BlueTM多功能细胞染色法测定阿霉素(ADR)对K562和K562/AO2细胞的半数抑制浓度(IC50)及判断CSA的非细胞毒性剂量;采用RT-PCR法检测GCS基因和mdr1基因的mRNA表达.结果 CsA浓度低于4.5μg/ml对K562/AO2细胞无细胞毒性.ADR对K562和K562/AO2细胞的IC50分别为(0.84±0.04)μg/ml和(89.24±1.27)μg/ml;当1μg/ml CsA和25μmol/L PPMP单用或联合作用于K562/AO2细胞时,ADR的IC50分别为(7.81±0.74)、(17.49±0.53)、(2.82±0.07)μg/ml;而低浓度CsA(0.01μg/ml)和PPMP(10 μmol/1)联合作用时IC50为(16.08±1.25)μg/ml.CsA联合PPMP可明显降低GCS基因的mRNA表达,低浓度CsA和PPMP联用也有此作用.结论 CsA联合PPMP可通过抑制K562/AO2细胞的GCS基因mRNA表达有效逆转其耐药,低浓度CsA联合PPMP在逆转其耐药的同时降低了细胞毒性.     

JAK2/STAT3信号转导通路参与调节IL-6介导的肺腺癌细胞株中MMP-10的表达

目的研究Janus激酶2（JAK2）及信号转导和转录激活子3（STAT3）途径对IL-6介导的肺腺癌细胞（A549）中基质金属蛋白酶10（MMP-10）表达调节的作用,探讨MMP-10的调节机制。方法待生长状况良好的A549细胞融合生长至50%～70%时,分别用0,12.5,25,50ng/mlIL-6及AG490（JAK2特异性抑制剂）＋25ng/mlIL-6处理A549细胞24h后,采用实时RT-PCR技术检测JAK2、STAT3和MMP-10mRNA的表达水平;应用Westernblot检测MMP-10蛋白的表达水平。结果25ng/mlIL-6组STAT3mRNA表达水平较0ng/ml组高43.87%（P〈0.05）,AG490＋25ng/mlIL-6组较25ng/mlIL-6组低36.73%（P〈0.01）;AG490＋25ng/mlIL-6组MMP-10mRNA表达水平较25ng/mlIL-6组高43.21%（P〈0.01）;12.5,25,50ng/mlIL-6组的MMP-10蛋白水平显著高于0ng/ml组（P〈0.05）,且峰值出现在25ng/ml组;与25ng/mlIL-6组相比,MMP-10蛋白水平在AG490＋25ng/mlIL-6组明显降低（P〈0.05）。结论JAK2/STAT3信号转导途径可能参与调节IL-6介导的肺癌细胞株A549中MMP-10的表达。