计量分析甘草甜素和绞股蓝总苷等对大鼠肝癌前病变的影响

采用被公认的以γ－谷氨酰转肽酶阳性灶（ＧＧＴ＋）为观察终点的二乙基亚硝胺（ＤＥＮ）致大鼠肝癌前病变的模型，对甘草甜素（ＧＬ）、纹股蓝总苷（绞股蓝总甙，ＧＰ）、茶多酚和柴胡水提取液等进行了筛选、并用体现学方法进行计量分析。结果显示，ＧＬ组的ＧＧＴ＋的面数由度（Ｎａ）为２４．１７±１．８３／ｃｍ２（ｘ±Ｓｘ，下同），体密度（Ｖｖ）为４．７４±０．５１％，数密度（Ｎｖ）为３９４．９±４５．２／ｃｍ３；ＧＰ组的Ｎａ为２５．３５±２．６４／ｃｍ２，Ｖｖ为５．９３±０．７５％，Ｎｖ为３７９．２±４８．４／ｃｍ２，这两组的３个估计值与对照组的３个估计值（Ｎａ为３７．２１±２．５６／ｃｍ２，Ｖｖ为８．１２±０．７０％。Ｎｖ为５７６．１±５８．０／ｃｍ３）分别比较时均有显著性差异，说明这两种药物对ＤＥＮ致肝癌发生有抑制作用，而茶多酚和柴胡水提取液则未显示抑制作用。     

胃癌7号染色体长臂的杂合性缺失分析

目的:检测胃癌患者7号染色体长臂微卫星位点的杂合性缺失(loss of heterozygosity,LOH),以初步确定7号染色体长臂上与胃癌相关基因连锁最密切的微卫星多态位点及LOH的临床意义.方法:在70例原发性胃癌中应用多重PCR技术扩增覆盖整个7号染色体长臂的9个微卫星位点(平均遗传距离为10cm),聚丙烯酰胺凝胶电泳分离PCR产物,用GeneScan、Genotyper软件进行分析.结果:9个微卫星位点的LOH均可发生于原发性胃癌,总的LOH频率为34.3%(24/70),其中D7S486和D7S798位点的LOH频率较高,分别为24.0%(12/50)和19.2%(5/26);总的LOH频率随临床分期而显著增高(P=0.046),D7S486位点的LOH频率在淋巴结转移者显著高于无淋巴结转移者(P=0.015).结论:在7号染色体长臂D7S486和D7S798位点附近,可能存在与胃癌发展相关的抑癌基因.     

细胞色素P450 CYP1A1基因多态与鼻咽癌易感性的关联分析

Objective To investigate the association between CYP1A1 gene polymorphisms and susceptibility of nasopharyngeal carcinoma in Cantonese nuclear families through family-based association study. Methods A total of 457 Cantonese nuclear families, consisting of 2134 members, were recruited as subjects. Each family included two parents and at least one offspring with nasopharyngeal carcinoma. Two single nucleotide polymorphisms (SNP) in CYP1A1 named m1 (rs4646903) and m2 (rs1048943) , were genotyped by PCR-RFLP assay and verified by directly sequencing. The genotype data were analyzed with family-based association test (FBAT) software to check the linkage and association between the two genetic markers and susceptibility of nasopharyngeal carcinoma. Results FBAT analysis showed that the minor allele frequencies (MAF) of the two SN P were 0. 442 (C) and 0. 339 (G) respectively. For m1 polymorphism in CYP1A1 gene was not significantly associated with nasopharyngeal carcinoma in our study population whether stratified with VCA-IgA or not (without stratification : X2=2. 399, P=0. 301 ; with stratification : Iow-titer group (VCA-IgA<1 : 80), MAF=0. 457 (C), X2=1.221, P=0.543 ; high-titer group (VCA-IgA ≥1 : 80), MAF=0. 427 (C), X2=2. 832, P=0. 243). For m2 polymorphism, when VCA-IgA<1 : 80, the G allele showed decreased transmission under additive and dominant model (MAF=0. 347 (G) ; Zadditive=-2. 120,Padditive=0. 034;Zdominant=-2. 303,Pdominant=0.021)and a boundary P value was got with global statistic (X2=5. 394, P=0. 067). Haplotype TG (0. 057), constructed by ml and m2, might decrease nasophargneal carcinoma risk (Z=-2. 002,P=0. 045). A boundary P value was also got with global statistic (X2=7. 067 ,P=0. 070). Conclusion There was no statistical significance between ml polymorphism and susceptibility of nasopharyngeal carcinoma in Cantonese nuclear families. And this study showed that m2 polymorphism might associated with the decrease of nasopharyngeal carcinoma in Cantonese nuclear families.     

鼻咽癌p53 p21 WAF1和MDM2蛋白异常表达的临床意义

目的：探讨p53、p21^WAF1、MDM2蛋白在原发鼻咽癌（NPC）异常表达的临床意义。方法：采用LSAB法检测69例原发NPC组织中p53、p21^WAF1和MDM2蛋白的表达状况。结果：1）p53、p21^WAF1和MDM2蛋白在原发NPC组织的阳性表达率分别为79．7％、84．1％、和82．6％；高表达率分别为50．7％、46．4％和31．9％。2）p53蛋白的高表达率随TNM分期的升高而增多，P＝0．042。3）p53或MDM2蛋白高表达者的复发间期显著短于相应蛋白低表达／阴性者，分别P＝0．038和P＝0．002。4）p53和MDM2蛋白同时高表达者、MDM2蛋白高表达同时p21^WAF1蛋白低表达／阴性者的复发间期明显短于对照组，分别P＜0．001；p21^WAF1蛋白高表达同时p53蛋白低表达／阴性者、p53和MDM2蛋白同时低表达／阴性者的复发间期显著长于对照组，分别P＝0．002和P＝0．014。5）p53和MDM2蛋白高表达同时p21^WAF1蛋白低表达／阴性者的复发间期明显短于对照组，P＜0．001；p53和MDM2蛋白低表达阴性同时p21^WAF1蛋白高表达者的复发间期明显长于对照组，P＝0．002。结论：p53或MDM2蛋白高表达提示有促进NPC复发的作用，p21^WAF1蛋白高表达提示有抑制NPC复发的作用。单独检测p53或MDM2蛋白在原发NPC组织的表达情况可以和为预测NPC复发倾向和临床预后的参考指标；如同时检测p53、MDM2和p21^WAF1蛋白的两项或三项指标，预测意义更理想。     

细胞色素P450 CYP1A1基因多态与鼻咽癌易感性的关联分析

目的 利用以核心家系为基础的关联研究探讨细胞色素P450酶系(cytochrome P450,CYP450)CYP1A1基因m1和m2多态性并分析其与鼻咽癌易感性的关联.方法 收集457个广东鼻咽癌核心家系(每个核心家系由患者和父母或同胞构成)共2134名成员作为研究对象,采用聚合酶链反应-限制性片段长度多态性(PCR-RFLP)方法对CYPlAl基因单核苷酸多态性(SNP)位点m1和m2(其参考编号分别为rs4646903和rs1048943)进行基因分型.挑选PCR产物测序验证.运用家系为基础的相关性检验(family-based association test,FBAT)软件分析这两个多态位点的基因型及其单体型与鼻咽癌易感性的关联.结果 FBAT软件分析结果 显示,位点m1和m2的微效等位基因频率(MAF)分别为0.442(C)和0.339(G).其中无论是否根据EB病毒壳抗原抗体(VCA-IgA)分层,m1位点与鼻咽癌的易感性之间关联均无统计学意义[未分层:X2=2.399,P=0.301;分层后:低滴度组(VCA-IgA<1:80),MAF=0.457(C),X2=1.221,P=0.543;高滴度组(VCA-IgA≥1:80),MAF=0.427(C),X2=2.832,P=0.243];对m2位点来说,末根据VCA-IgA分层时,该位点与鼻咽癌的易感性之间关联无统计学意义(X2=2.694,P=0.260).分层后,在低滴度组,累加和显性模式下,位点m2等位基因G显示了从亲代到子代传递减少[MAF=0.347(G);Z<,累加>=-2.120,P<,累加>=0.034;Z显性=-2.303,P显性=0.021],全局统计也提示了传递的改变(X2=5.394,P=0.067);由这两个位点构建的单体型TG(0.057)可能降低鼻咽癌的发病风险(Z=-2.002,P=0.045),全局统计也提示CYP1A1基因单体型可能与鼻咽癌易感性有关系(X2=7.067,P=0.070).结论 以家系为基础的相关性研究发现CYP1A1基因多态位点m1与鼻咽癌的易感性之间关联无统计学意义,位点m2基因多态性可能与鼻咽癌的发病风险降低有关系.     

细胞色素P450 CYP1A1基因多态与鼻咽癌易感性的关联分析

Objective To investigate the association between CYP1A1 gene polymorphisms and susceptibility of nasopharyngeal carcinoma in Cantonese nuclear families through family-based association study. Methods A total of 457 Cantonese nuclear families, consisting of 2134 members, were recruited as subjects. Each family included two parents and at least one offspring with nasopharyngeal carcinoma. Two single nucleotide polymorphisms (SNP) in CYP1A1 named m1 (rs4646903) and m2 (rs1048943) , were genotyped by PCR-RFLP assay and verified by directly sequencing. The genotype data were analyzed with family-based association test (FBAT) software to check the linkage and association between the two genetic markers and susceptibility of nasopharyngeal carcinoma. Results FBAT analysis showed that the minor allele frequencies (MAF) of the two SN P were 0. 442 (C) and 0. 339 (G) respectively. For m1 polymorphism in CYP1A1 gene was not significantly associated with nasopharyngeal carcinoma in our study population whether stratified with VCA-IgA or not (without stratification : X2=2. 399, P=0. 301 ; with stratification : Iow-titer group (VCA-IgA<1 : 80), MAF=0. 457 (C), X2=1.221, P=0.543 ; high-titer group (VCA-IgA ≥1 : 80), MAF=0. 427 (C), X2=2. 832, P=0. 243). For m2 polymorphism, when VCA-IgA<1 : 80, the G allele showed decreased transmission under additive and dominant model (MAF=0. 347 (G) ; Zadditive=-2. 120,Padditive=0. 034;Zdominant=-2. 303,Pdominant=0.021)and a boundary P value was got with global statistic (X2=5. 394, P=0. 067). Haplotype TG (0. 057), constructed by ml and m2, might decrease nasophargneal carcinoma risk (Z=-2. 002,P=0. 045). A boundary P value was also got with global statistic (X2=7. 067 ,P=0. 070). Conclusion There was no statistical significance between ml polymorphism and susceptibility of nasopharyngeal carcinoma in Cantonese nuclear families. And this study showed that m2 polymorphism might associated with the decrease of nasopharyngeal carcinoma in Cantonese nuclear families.     

化疗药物对肿瘤微血管的毒性作用

一般认为，化疗药物是非特异的血管生成抑制剂。如果部分化疗药物在体内有较强的血管毒性，在常规化疗中恰当应用该类药物有可能改善化疗反应，提高实体瘤缓解率.     

肝癌体外药物敏感性试验

目的 通过测定肝癌对阿霉素（ADM）、顺铂（DDP）、氟尿嘧啶（FU）、长春新碱（VCR）、丝裂霉素C（MMC）及噻替哌（TSPA）的体外药物敏感性，为肝癌化疗用药提供参考依据。方法 采用滤纸支持组织块培养－MTT终点－计算机图像分析体外药物敏感性试验法。结果 在1倍血浆峰浓度（1 PPC）时作用最强的药物为MMC，其抑制率中位数（MIR）为26．5％，其次是ADM为13．0％，DDP、TSPA、5－FU和VCR为4．0－8．5％。在5倍血浆峰浓度（5PPC）时作用最强的药物亦为MMC，其MIR为92．0％，次为ADM、DDP和TSPA为41－50％，5－FU和VCR为16．0－17．0％，DDP和TSPA在5PPC的MIR比在1PPC时大5．0和5．8倍以上，MMC、ADM帮VCR则大2－3．5倍，5－FU只增大1倍左右。结论 结果提示MMC对肝癌抑制作用较强，DDP和TSPA剂量增加与疗效提高可能有较大的关系。天然来源的药物ADM、MMC和VCR之间，无论在1PPC或5PPC水平，其相关系数均较大（r=0.5539-0.7208)，表明部分肝癌具有多药耐药性。     

细胞色素P450 CYP1A1基因多态与鼻咽癌易感性的关联分析

Objective To investigate the association between CYP1A1 gene polymorphisms and susceptibility of nasopharyngeal carcinoma in Cantonese nuclear families through family-based association study. Methods A total of 457 Cantonese nuclear families, consisting of 2134 members, were recruited as subjects. Each family included two parents and at least one offspring with nasopharyngeal carcinoma. Two single nucleotide polymorphisms (SNP) in CYP1A1 named m1 (rs4646903) and m2 (rs1048943) , were genotyped by PCR-RFLP assay and verified by directly sequencing. The genotype data were analyzed with family-based association test (FBAT) software to check the linkage and association between the two genetic markers and susceptibility of nasopharyngeal carcinoma. Results FBAT analysis showed that the minor allele frequencies (MAF) of the two SN P were 0. 442 (C) and 0. 339 (G) respectively. For m1 polymorphism in CYP1A1 gene was not significantly associated with nasopharyngeal carcinoma in our study population whether stratified with VCA-IgA or not (without stratification : X2=2. 399, P=0. 301 ; with stratification : Iow-titer group (VCA-IgA<1 : 80), MAF=0. 457 (C), X2=1.221, P=0.543 ; high-titer group (VCA-IgA ≥1 : 80), MAF=0. 427 (C), X2=2. 832, P=0. 243). For m2 polymorphism, when VCA-IgA<1 : 80, the G allele showed decreased transmission under additive and dominant model (MAF=0. 347 (G) ; Zadditive=-2. 120,Padditive=0. 034;Zdominant=-2. 303,Pdominant=0.021)and a boundary P value was got with global statistic (X2=5. 394, P=0. 067). Haplotype TG (0. 057), constructed by ml and m2, might decrease nasophargneal carcinoma risk (Z=-2. 002,P=0. 045). A boundary P value was also got with global statistic (X2=7. 067 ,P=0. 070). Conclusion There was no statistical significance between ml polymorphism and susceptibility of nasopharyngeal carcinoma in Cantonese nuclear families. And this study showed that m2 polymorphism might associated with the decrease of nasopharyngeal carcinoma in Cantonese nuclear families.     

原发性肝细胞癌RIT1基因的突变和扩增

目的　探讨 RIT1基因突变和扩增在肝细胞癌 (hepatocellular carcinoma,HCC)的发生情况及其与发病的关系。　方法　采用 PCR直接测序法检测 5 0例原发性 HCC患者的肝癌组织和非癌肝组织RIT1基因在所包含的 6个外显子的全序列寻找突变位点 ;并用荧光定量 PCR法检测 RIT1基因的扩增情况。　结果　在 5 0例肝癌组织中 1例出现第 5外显子编码区 2 4 1位核苷酸 G/ C变异 ,其对应密码子改变为GAG81CAG,编码氨基酸改变为 Glu81Gln,该氨基酸变异位于 GTP结合的保守功能域内 ,该病例的非癌肝组织以及其余 4 9例的肝癌组织和非癌肝组织均未发生此种改变 ;在 5 0例肝癌组织和非癌肝组织中均出现 5′- UTR(起始密码子前 2 1位核苷酸 ) G/ C变异 ;在获得有效扩增数据的 4 3例肝细胞癌患者中 ,11例有 2～2 97倍的 RIT1基因扩增 ,扩增率为 2 5 .6 %。　结论　基因扩增是 RIT1基因在肝细胞癌的激活方式之一 ,可能与肝细胞癌的发病有关 ,而点突变方式可能意义不大。