细胞分裂周期蛋白73基因缺失抑制粟酒裂殖酵母细胞的有性生殖和有丝分裂

cdc73基因编码粟酒裂殖酵母(Schizosaccharomyces Pombe)RNA聚合酶Ⅱ辅助因子Cdc73,参与G_(2)期检查点激活并调控细胞周期。但cdc73基因缺失对细胞有丝分裂动力学的调控尚不清楚。本研究采用活细胞成像、荧光蛋白标记技术探究粟酒裂殖酵母cdc73基因缺失后对细胞有性生殖和细胞有丝分裂中微管、肌动蛋白、线粒体和组蛋白动力学的影响。结果表明:在有性生殖中,cdc73基因缺失会导致子囊孢子长度增加14.23%,产4个孢子数量的细胞减少64.08%;活细胞成像结果分析发现,在有丝分裂中,分裂后期微管伸长的长度缩短11.21%,伸长时间减少17.39%;肌动蛋白环形成和收缩速率分别降低33.33%和26.09%,形成和收缩时间分别延长58.00%和40.38%;同时,肌动蛋白环、线粒体和组蛋白表达量都增加。本研究揭示了cdc73基因缺失可抑制有丝分裂中纺锤体的伸长,延缓肌动蛋白环的形成与收缩,为进一步探寻Cdc73蛋白在细胞分裂中参与调控微管和肌动蛋白动力学功能提供了一定的科学依据。

水稻OsBP-73基因表达需要其内含子参与

该实验室以前的研究表明,水稻OsBP-73基因含有2个外显子和1个长度为2 471 bp的内含子。该文报告用OsBP-73基因ATG翻译起始密码子(在第1外显子中)上游序列 (-1 818^+215)与GUS基因构成嵌合质粒pRSS1,将该质粒转化水稻后,在抗性愈伤组织和转基因植株中未能检测到GUS基因的表达。只有用含有完整的内含子及其上游序列(-1 818^+2 844)与GUS基因构成嵌合质粒(p13GNF)时,才能在p13GNF的转基因抗性愈伤组织和植株中检测到GUS基因的表达。实验还证明,单是内含子序列并不能驱动GUS基因在转基因水稻中表达。由此推测:OsBP-73基因的启动子序列驱动基因表达时,需要基因内含子的参与。

水稻转录因子OsBP-73靶基因的初步筛选

OsBP-73是用酵母单杂交系统,以水稻蜡质基因(Wx)的顺式作用元件为诱饵,从水稻cDNA表达文库中筛选获得的转录因子。本文以OsBP-73基因为例介绍了用"下拉"(pull-down)方法筛选转录因子靶基因的一般步骤。将含有该基因DNA结合功能域的cDNA片段构建到原核表达载体上,并在大肠杆菌中诱导表达获得其蛋白p73。用纯化后的p73蛋白通过"下拉"实验对OsBP-73靶基因进行初步筛选,获得了22个阳性克隆,为进一步研究转录因子OsBP-73参与的水稻转录调控网络提供依据。

Expression of SNC73, a transcript of the immunoglobulin α-1 gene, in human epithelial carcinomas

AIM： To investigate the expression of SNC73, a transcript of the immunoglobulin α-1 gene （IgA1-H chain）, in human epitheliα-derived tumor cells. METHODS： Total RNAs and cell lysates were prepared from five different human epithelial cell lines derived from lung, stomach, liver, skin, and breast, respectively. RT-PCR and immunoblot analysis of these five cell lines were done. Both RT-PCR and immunochemistry were used to detect the expression of SNC73 in these cell lines. We also examined the expression of SNC73 in normal epithelial cells of colon mucosa by in situ hybridization. RT-PCR and immunoblot analysis were used to determine whether the recombination activating gene1/2 （RAG1 and RAG2） is present. The expression of three immunoglobulin transcription factors, EBF, E2A and Pax5, and the heavy chain of IgA1 and two types of light chains of immunoglobulin （κ and λ） in the aforementioned cell lines were analyzed by RT-PCR and immunochemistry, respectively. All the RT-PCR products were analyzed by sequencing. RESULTS： The results of RT-PCR and immunochemistry showed that both mRNA and protein of SNC73 were expressed in five human epitheliα-derived cancer cell lines. These data were further confirmed in the normal epithelial cells of colon mucosa by in situ hybridization. Also, the heavy chain of IgA1 and κ light chain were detected in these cells, but no λ light chain was observed. Both RAG1 and RAG2 were expressed in these human epitheliα-derived cancer cell lines and the sequence was identical to that expressed in pre-B and pre-T cells. In addition to RAG1 and RAG2, the mRNA in one of the immunoglobulin transcription factors, EBF, was also detected in these cell lines, and Pax5 was only expressed in SW480 cells, but no expression of E2A was observed in all the five cell lines. CONCLUSION： Immunoglobulin A1 is originally expressed and V（D）J recombination machine is also present in non-lymphoid cells, suggesting that V（D）J recombination machine mediates the assembly of immunoglobulin A1 in non-lymphoid cells as in prelymphocytes.

Enhanced chemosensitivity of p73α gene transferred into H1299 cell line of human lung adenocarcinoma

Objective: To study the effects of transferred wild type p73α gene on the sensitivity to the chemotherapeutic agents and the growth of p53-null H1299 cells of human lung adenocarcinoma. Methods: The pcDNA3-HA-p73α plasmid was transferred into the cultured p53-null H1299 cells of human lung adenocarcinoma with the mediation of Dosper liposome; The cells resistant to G418 were selected. The expression of p73α gene in the cells was examined with Western blot. MTT assay was used to analyze the response of the transfected cells to cis-dichlorodiamine platinum (cDDP) and adriamycin (ADM). The rate of drug-induced apoptosis of the transfected cells was determined with flow cytometry and DNA fragmentation assay. The changes of the biological behaviors were observed with colony formation assay. Results: The transfected H1299 cells of human lung adenocarcinoma over-expressed p73α protein stably. MTT assay showed that the IC 50 values of cDDP and ADM were reduced by approximately 7 fold and 130 fold respectively in the transfected cells as compared with the untransfected ones. Lower concentration of the chemotherapeutic agents (1.25 μmol/L of cDDP and 0.05 μmol/L of ADM) could be employed to suppress markedly the growth of the transfected H1299 cells. The apoptotic rate induced by cDDP was increased from 10.1% to 38 4% (P<0.01) and that of ADM from 12.1% to 49.3% (P<0.01). The clonogenecity after the administration of chemotherapeutic agents was significantly lower in the transfected H1299 cells than in the parental cells (P<0.01). The sensitive enhancement ratios were 1.8 and 2.6 for cDDP and ADM respectively. Conclusion: The transfection of H1299 cells with wild type p73α gene results in an increase of the sensitivity of the cells to chemotherapeutic agents.

Association of the p73 Gene G4C14-to-A4T14 Polymorphism with Increased Gastric Cancer Risk in a Northwestern Chinese Population

OBJECTIVE To evaluate the p73 gene G4C14-to-A4T14 double nucleotide polymorphism with both increased gastric cancer(GC) risk and different histological subtypes of GC in a northwestern Chinese population. METHODS Genotyping of the polymorphism of the p73 gene was conducted with PCR-CTPP. RESULTS All 385 GC patients including 305 diffuse-type and 80 intestinal-type cases and 412 healthy controls were investigated.The frequencies of p73 AT/AT,AT/GC,and GC/GC genotypes were 28.1%,47.1%,and 24.8% in the controls,and were 22.0%,45.0%,and 33.0% in GC cases respectively;the GC/GC homozygote frequency was higher in GC cases,mainly in diffuse type compared to the controls with OR=1.71(1.16～2.51) and 1.87 (95%CI,1.24～2.81) respectively.The results showed that carriers of the p73 G4A GC/GC homozygote had a 1.71-time higher risk of GC,especially of the diffuse-type GC compared to the controls. The carriers of the AT/GC heterozygote also had a slightly increased risk of GC cancer,mainly on intestinal-type GC.This is the first report that the p73 G4A double-nucleotide polymorphism is associated with an increased risk of diffuse-type gastric cancer. CONCLUTION The p73 G4A GC/GC genotype is associated with an increased risk of gastric cancer,especially of the GC diffuse-type.

基于CRISPR/Cas9技术构建GP73基因敲除小鼠模型及表型验证

目的:基于CRISPR/Cas9技术构建高尔基体73(GP73)基因敲除小鼠模型并对其表型进行初步验证。方法:利用非同源末端连接(NHEJ)修复引入突变的方式,造成GP73基因蛋白读码框移码,使其功能缺失,针对GP73基因外显子4设计sgRNA靶位点,体外转录获得sgRNA,与编码Cas9的mRNA混合后通过受精卵显微注射方法获得F0代阳性敲除小鼠,通过繁育及基因型鉴定获得GP73基因敲除纯合子小鼠(GP73-/-小鼠)。采用实时荧光定量PCR(RT-qPCR)法检测肝、肾、皮下脂肪、棕色脂肪组织中GP73 mRNA表达,采用酶联免疫吸附试验(ELISA)法检测血清GP73蛋白水平。结果:与野生型(WT)小鼠相比,GP73-/-小鼠肝、肾、皮下脂肪、棕色脂肪组织GP73 mRNA及蛋白表达水平降低,血清GP73蛋白水平降低(均P<0.05)。结论:基于CRISPR/Cas9技术成功构建了GP73基因敲除小鼠。

血清GP73、HMGB1、FLP1、sST2与慢性乙型病毒性肝炎患者 HBV-DNA载量、肝功能及肝纤维化标志物的相关性分析

目的 研究血清高尔基体蛋白73(GP73)、高迁移率族蛋白B1(HMGB1)、纤维蛋白原样蛋白1(FLP1)、可溶性生长刺激表达基因2蛋白(sST2)与慢性乙型病毒性肝炎(CHB)患者乙型肝炎病毒-脱氧核糖核酸(HBV-DNA)载量、肝功能指标及肝纤维化标志物的相关性。方法 选择滨州市中医医院2021年2月至2022年2月收治的166例CHB患者作为研究对象。测定所有患者的HBV-DNA载量,并根据HBV-DNA载量的差异分为低载量组、中载量组、高载量组。另选取同期健康体检人员60例作为健康对照组。检测并比较各组肝功能指标水平、肝纤维化标志物水平,以及血清GP73、HMGB1、FLP1、sST2水平。以Spearman/Pearson相关分析血清GP73、HMGB1、FLP1、sST2与HBV-DNA载量、肝功能指标及肝纤维化标志物的相关性。结果 低载量组76例,中载量组50例,高载量组40例。低载量组、中载量组、高载量组血清GP73、HMGB1及sST2水平均高于健康对照组(P<0.05);且随着HBV-DNA载量的增加,GP73、HMGB1及sST2水平升高(P<0.05)。低载量组、中载量组、高载量组血清FLP1水平均低于健康对照组(P<0.05);且随着HBV-DNA载量的增加,FLP1水平下降(P<0.05)。中载量组、高载量组丙氨酸氨基转移酶(ALT)、天冬氨酸氨基转移酶(AST)及γ-谷氨酰转移酶(GGT)水平均高于低载量组(P<0.05),且高载量组ALT、AST及GGT水平高于中载量组(P<0.05)。中载量组、高载量组透明质酸(HA)、层粘连蛋白(LN)及Ⅳ型胶原(CⅣ)水平均高于低载量组(P<0.05),且高载量组HA、LN及CⅣ水平均高于中载量组(P<0.05)。血清GP73、HMGB1、sST2与CHB患者HBV-DNA载量、ALT、AST、GGT、HA、LN、CⅣ水平均呈正相关(r>0,P<0.05),而血清FLP1与CHB患者HBV-DNA载量、ALT、AST、GGT、HA、LN、CⅣ水平呈负相关(r<0,P<0.05)。结论 血清GP73、HMGB1、FLP1、sST2水平可有效反映CHB患者的HBV-DNA载量、肝功能和肝纤维化情况。

小桐子过氧化物酶73基因的克隆及表达分析

过氧化物酶73属于植物特异Ⅲ型过氧化物酶家族成员,通过消除活性氧、酚类及胺类等毒害作用参与植物多种抗逆性形成过程。为探索小桐子POD73基因对低温环境的响应机制,本研究基于小桐子低温锻炼转录组数据,以茎为材料,采用RT-PCR技术克隆到小桐子过氧化物酶73基因(Jc POD73)的全长c DNA序列。结果表明,该c DNA全长1 516 bp,含有完整的开放阅读框(987 bp),编码328个氨基酸,分子量为35.8 k Da,理论等电点为9.16。其推导的氨基酸序列及空间结构,包含该家族典型的2个Ca2+结合基序以及血红素活性中心。半定量RT-PCR表达分析显示,Jc POD73在小桐子各组织中都有表达,但表达水平具有组织特异性,其中在根与茎中表达量较高,且受低温诱导表达显著,而在叶中表达量相对较低。本研究为进一步阐明小桐子POD73基因的功能及其在小桐子遗传改良中的应用奠定了基础。

新基因SNC66和SNC73在大肠粘膜和大肠癌中表达的比较

目的 :研究新基因SNC 6 6和SNC 73的表达状况 ,并探讨其与大肠癌发生的相关性。方法 :采用体外转录的方法合成地高辛标记的探针 ,并用它进行cRNA/mRNA原位分子杂交 ,检测37例大肠癌标本及其相应正常大肠粘膜中SNC 6 6和SNC 73两基因表达mRNA的状况 ,并加以比较。结果 :SNC 6 6和SNC 73基因只在上皮细胞和淋巴细胞中表达 ,在大肠癌组织中都存在明显的表达缺陷 ,尤其以SNC 73的表达缺陷更明显。SNC 6 6在粘膜绒毛上多呈梯度性表达 ,以隐窝处最深 ,到绒毛顶端逐渐变淡 ;SNC 73则多呈均一性表达。结论 :SNC 6 6和SNC 73是表达行为、代谢途径均有所不同的两个免疫球蛋白样大肠癌负相关基因 。

基因重组人HSC73在大肠杆菌中的表达和纯化

目的 用基因工程方法表达人的热休克类似蛋白 73 ( HSC73 )并提纯 HSC73蛋白 ,为研究该蛋白提供条件。方法 用人 HSC73基因构建原核细胞表达载体 p ETHSC73 ,在大肠杆菌 BL 2 1中用异丙基硫代 -β-D半乳糖苷 ( IPTG)诱导表达 ,ATP亲和层析法提取 HSC73。经 SDS-PAGE、3 .a.3抗体 Western blot作 ECL检测。结果 人HSC73基因构建的载体 p ETHSC73能很好地在大肠杆菌表达出分子量为 73 k D的蛋白 ,蛋白提取方法能有效提纯表达的蛋白 ,用 3 .a.3抗体作 Western blot检测证实该蛋白具有人 HSC73抗原特性。所得蛋白质氨基酸测定和 N端测序的结果与有关报道的一致。结论 人 2 .1kb的 HSC73基因片段构建的高表达载体能够很好表达人 HSC73 ,用 ATP亲和层析法可以有效提纯有活性的人

基因重组人HSP73抗体的制备及对肿瘤细胞在温热作用后HSP70表达的检测

目的 用基因工程方法表达、提纯人热休克类似蛋白 73(HSP73) ,制备抗体 ,检测吐温 80协同 41℃温热对肿瘤细胞HSP70表达的影响。方法 用人HSP73基因构建原核细胞表达载体 pETHSP73,在大肠杆菌BL2 1中用半乳糖苷 (IPTG)诱导表达 ,经电泳法和ATP亲和层析法提取HSP73并作检测。用提纯蛋白免疫新西兰兔 ,制备抗HSP70多克隆抗体 ,测定其活性 ,用此抗体作免疫组化法观察 41℃温热和吐温 80协同时BGC 82 3胃癌细胞及B16黑色素瘤细胞的热休克蛋白 70 (HSP70 )表达情况。结果 人基因重组构建的pETHSP73能很好地在大肠杆菌中表达出分子量为73kD的蛋白。两种提纯方法均能有效提取表达蛋白 ,用 3a3单抗作Westrnblot检测证明该蛋白具有人HSP70的抗原活性 ,所得蛋白氨基酸测定和N端测序的结果与有关报道一致。以此蛋白制成的抗体活性可达 1∶80 0 0以上。并可检测出吐温 80合并温热有明显抑制肿瘤细胞SHP70表达的作用。结论 2 .1kb的基因片段构建的高表达载体能很好表达人HSP73活性蛋白。该蛋白免疫新西兰兔所得多克隆抗体具有很高抗人和鼠HSP70活性。用该抗体免疫组化法检测表明吐温 80协同 41℃温热可明显抑制肿瘤细胞HSP70的表达和耐热性的产生。

‘烟73’葡萄果皮花色苷合成、修饰与转运相关基因的表达

花色苷在内质网中合成,而在液泡中发挥生理学功能,因此花色苷液泡转运过程非常关键。研究葡萄果皮中花色苷合成,修饰与转运基因的表达模式,有助于解析花色苷从合成到转运的完整通路。以宁夏青铜峡地区红色酿酒葡萄品种‘烟73’为材料,在果实成熟过程中测定可溶性固形物、可滴定酸含量pH及果皮总酚、单宁、花色苷含量,利用实时荧光定量PCR检测葡萄果皮中花色苷合成,修饰与转运相关基因表达水平。结果表明,成熟期‘烟73’可溶性固形物、可滴定酸和pH分别为(23.50±0.30)%、(3.22±0.03)g·L-1和3.59±0.10;果皮单宁、总酚及花色苷含量分别为(22.32±1.15)mg·g^-1(果皮干质量)、(62.50±1.23)mg·g^-1和(52.49±1.10)mg·g^-1。定量PCR结果显示,在果实成熟过程中,‘烟73’葡萄果皮VviUFGT、VviGST1、VviGST2、VviGST3及VviGST4基因表达趋势基本相同,呈先上升后下降趋势,且在花后10周表达量达最高值;VviOMT基因表达总体呈下降趋势,花后8周和10周同其他时期相比存在显著性差异;AM1与AM3基因表达呈“M”型变化趋势,花后10周与14周表达较高。相关性分析显示,‘烟73’中VviGST1、VviGST2、VviGST3以及VviGST4基因的表达趋势与花色苷积累量显著正相关,VviGST1与VviOMT显著正相关。综上所述,‘烟73’葡萄果实理化及多酚成熟度较好,4个GST家族基因均参与了花色苷的液泡转运,其中VviGST1可能参与了‘烟73’果皮中甲基化花色苷的转运。

High expression of △Np73 predicts poor prognosis in NSCLC cancer patients

Objective: To study the expression of △Np73, an isoform of the p53 homologue p73, in the different stages of human non-small-cell lung cancer (NSCLC) and the association of△Np73 expression with in patient survival. Methods: Semi-quantitative reverse transcriptase polymerase chain reaction (RT-PCR) was used to study the expression of△Np73 mRNA in 51 resected NSCLC tissues. Its relation to clinicopathological factors and survival outcome were analyzed. Results: The positive rate and expression level of△Np73 mRNA in the cancer tissues were significantly higher than that in the matched non-cancer lung tissues. The incidence of positive expression of△Np73 was 50. 0% , 52. 6% , and 87. 5% in patients with stageⅠ,Ⅱ, andⅢ, respectively. Positive expression of△Np73 was associated with pathological TNM stage (P = 0. 046), while not with age, gender, histological type and differentiation status. Survival rate of patients with high△Np73 mRNA was significantly poorer than those with low△Np73 mRNA levels (P<0. 001). Multivariate analysis revealed that△Np73 mRNA levels were a significant prognostic factor, independent of the other conventional prognostic factors. Conclusion: NSCLC has over-expression of△Np73 mRNA, which is closely related to TNM stages and prognosis of patients with NSCLC. These results suggest that measurement of△Np73 mRNA levels in tumor tissues might be useful as a promising predictor for the prognosis of patients with NSCLC.

抑癌基因P^(73)研究近况

P73 基因是Kaghad等在 1997年克隆得到的一个新基因 ,与P53 具有相似的结构、功能与活性 ,但其是否作为一个抑癌基因发挥作用 ,目前尚存在争论。本文就P73 的发现定位、结构、功能及其与肿瘤发生发展的关系作一综述。

5型腺病毒E1A基因通过调控GP73的表达抑制肝癌细胞增殖的初步研究

目的:研究5型腺病毒(Ad5)E1A基因通过调控高尔基蛋白73(GP73)的表达水平,抑制肝癌细胞HepG2的增殖。方法:以腺病毒为模板、pcDNA-flag-Vector为载体,构建真核表达载体pcDNA3-Flag-Ad5E1A;免疫印迹实验鉴定重组蛋白Ad5E1A的表达;免疫印迹实验验证Ad5E1A对GP73表达水平的影响;将Ad5E1A基因转染HepG2细胞,用CCK-8试剂盒检测HepG2细胞的增殖情况。结果:免疫印迹实验结果证实,真核表达载体pcDNA3-Flag-Ad5E1A在HEK293细胞中得到表达;Ad5E1A可明显抑制GP73的表达。酶标仪检测发现,与转染Flag-GP73组相比,Ad5E1A组的抑制率为17.5%,差异无统计学意义(P>0.05);而与转染Flag-GP73组比,共转Ad5E1A和GP73组的抑制率为37.8%,差异有统计学意义(P<0.05)。结论:Ad5E1A基因通过调控GP73的表达抑制了肝癌细胞HepG2的增殖。为研究肝癌发生机制,开发新型治疗方案提供了实验基础。

检测高尔基体蛋白73及其基因表达对原发性肝癌的诊断价值

目的：探讨高尔基体蛋白73（GP73）血清及其基因表达在原发性肝癌（PHC）中的诊断价值,分析GP73在PHC诊断及高危人群普查的临床意义。方法：采用酶联免疫吸附试验（ELISA）和电化学发光免疫技术定量检测85例PHC、46例肝硬变患者和102名健康人的血清GP73、AFP水平,计算GP73的敏感度、特异度和受试者工作特征曲线（ROC曲线）下面积,采用实时荧光定量（RT-PCR）法检测各组外周血单个核细胞GP73 mRNA的相对表达量,以Ct值比较法计算GP73 mRNA相对表达水平,同时检测12份正常肝组织和12份肝癌组织的GP73 mRNA相对表达水平。结果：PHC组和肝硬变组患者的GP73中位血清质量分数分别为295.6μg/L、213.9μg/L,PHC组GP73中位血清质量分数较肝硬变组明显升高,有显著性差异（P〈0.05）。PHC组和肝硬变组患者的AFP中位血清质量分数分别为9.6μg/L、7.0μg/L,无显著性差异（P〉0.05）。GP73诊断肝癌的敏感度（72.7%）高于AFP（49.8%）,有显著性差异（P〈0.05）,GP73特异度（70.8%）低于AFP（95.9%）,有显著性差异（P〈0.05）。GP73和AFP联合检测肝癌的敏感度为79.6%,特异度为80.3%,ROC曲线下面积为0.840（95%CI为0.770~0.919）;全血GP73 mRNA含量3组间无显著性差异（P〉0.05）,肝癌组织GP73 mRNA表达量（12.87）明显高于正常肝组织（1.08）,有显著性差异（P〈0.05）。结论：GP73诊断PHC具有较好的敏感度,GP73与AFP联合检测可进一步提高诊断早期肝癌的敏感度。全血标本GP73 mRNA检测不能作为诊断原发性肝癌的肿瘤标志,肝组织标本GP73 mRNA可为诊断原发性肝癌的肿瘤标志,但存在创伤大、风险大等弊端。血清GP73联合AFP检测可显著提高PHC诊断,二者联合可用于PHC高危人群的普查及筛选。

Overexpression of a Glycosyltransferase Gene from a Metabolically Poly-Resistant Beckmannia syzigachne Population Alters Growth and Confers Herbicide Resistance to Brachypodium distachyon

Beckmannia syzigachne is a noxious weed for rice-wheat rotations in China.The B.syzigachne(AH-02)population evolved metabolic resistance to fenoxaprop-P-ethyl and mesosulfuron-methyl.To investigate the function of GT73C1 in this population,the GT73C1 gene was amplified by reverse transcription-polymerase chain reaction,and the sequence was 100%consistent with the transcriptome data.Its phylogenetic tree was displayed and annotated using FigTree v1.4.4.The plant overexpression vector of GT73C1 gene was constructed and used to transform Brachypodium distachyon plants.Furthermore,the expression of GT73C1 was significantly induced by fenoxaprop-P-ethyl and mesosulfuron-methyl,which was consistent with the findings from the whole plant bioassay.These results indicate that GT73C1 is closely related to the metabolic resistance of B.distachyon.

肿瘤标志物对原发性肝癌的诊断意义

原发性肝癌(PHC)是常见的恶性肿瘤之一,早期不易发现,晚期的治疗效果较差,病死率很高。因此,早发现、早诊断、早治疗就成为延长患者生存期、降低病死率的关键。甲胎蛋白(AFP)是诊断PHC公认的肿瘤标志物,但存在较高的假阴性,为弥补其实验室诊断的不足、提高PHC的检出率,高尔基体蛋白73、DLC-1基因、α-L-岩藻糖苷酶、米托蒽醌抗性基因、磷脂酰肌醇蛋白多糖3和转化生长因子β1等特异性、敏感性更高的肿瘤标志物已逐渐被应用于临床。

原发性肝细胞癌患者p53基因突变的特征和临床意义

目的 探讨p53基因突变在原发性肝细胞癌(hepatocellular carcinoma, HCC)患者中的特征和临床意义,并分析其对预后的影响。方法 选取我院2019年1月1日—2020年4月30日收治的慢性乙型肝炎肝硬化和HCC患者各120例,分别作为慢性乙型肝炎肝硬化组和HCC组。选取同期于我院行体检的120例健康体检者作为健康对照组。采用化学发光法检测HCC患者血清特异性肿瘤标志物甲胎蛋白异质体(alpha-fetoprotein L3, AFP-L3)、高尔基糖蛋白-73(golgi protein 73, GP73)、异常凝血酶原(des-γ-carboxy-prothrombin, DCP)水平。采用聚合酶链反应-限制性片段长度多态性方法检测3组p53第4外显子72位密码子(p53 Arg72Pro)基因型,分析不同组间p53 Arg72Pro基因型差异,并比较HCC组患者不同p53 Arg72Pro基因型间AFP-L3、GP73、DCP水平及HBV DNA阳性率和生存率差异。结果HCC组携带Pro/Pro基因型患者比例为33.3%,高于慢性乙型肝炎肝硬化组的14.2%和健康对照组的13.3%,差异均具有统计学意义(P均<0.05)。在HCC患者中,Pro/Pro型患者发病年龄明显小于Arg/Pro型和Arg/Arg型患者,而巴塞罗那分期为C/D期和伴有淋巴结转移的患者比例明显高于Arg/Pro型和Arg/Arg型患者,HBV DNA检测阳性率明显高于Arg/Arg型,血清特异性肿瘤标志物AFP-L3、GP73、DCP水平均明显高于Arg/Arg型和Arg/Pro型患者,差异均具有统计学意义(P均<0.05)。Arg/Arg型HCC患者生存率为83.3%,高于Arg/Pro型和Pro/Pro型患者的63.3%,差异具有统计学意义(P <0.05)。结论 p53 Arg72Pro基因突变与HCC的发生存在密切关联,且与多个HCC血清特异性肿瘤标志物密切相关,可能是加速患者癌变进展过程中的关键调节因子。