人SOX7启动子荧光素酶报告质粒构建及HMGA2对其活性影响的研究

目的 研究发现,HMGA2促进肿瘤细胞发生EMT与其下游基因SOX7有关.为进一步验证转录因子HM-GA2与其下游基因SOX7之间的调控关系,本研究通过构建人SOX7基因启动子荧光素酶报告质粒,采用荧光素酶报告实验观察HMGA2对SOX7基因启动子荧光素酶活性的影响,为研究HMGA2转录表达的调控机制提供必要的实验基础.方法 采用PCR技术扩增人SOX7基因启动子序列(-1 549～+79nt),将SOX7基因启动子插入荧光素酶报告基因载体pGL3-basic中构建质粒pGL3-SOX7-promoter;再分别将pGL3-SOX7-promoter、pGL3-basic-SOX7-promoter和对照质粒(pLV-UbC-IRES2-EGFP)、pGL3-SOX7-promoter和HMGA2表达质粒(pLV-UbC-HMGA2-IRES2-EGFP)3组质粒共转染293T细胞,培养48 h后,检测3组荧光素酶的表达情况,观察HMGA2对SOX7基因启动子的调控作用.结果 通过菌落PCR和核酸测序证实,人SOX7基因启动子荧光素酶报告质粒pGL3-SOX7-promoter构建成功;将pGL3-SOX7-promoter和HMGA2表达质粒(pLV-UbC-HMGA2-IRES2-EGFP)共转染293T细胞48 h后,与对照组相比,SOX7基因启动子介导荧光素酶的活性降低约31％,受到明显抑制(P=0.003),提示HMGA2可能调控SOX7基因启动子的活性.结论 本研究成功构建了SOX7基因启动子荧光素酶报告质粒,初步验证了SOX7是HMGA2下游的作用靶点.     

E2F1对XRCC1启动子调节作用的研究

目的:探讨E2F1对XRCC1启动子的调节作用及其意义.方法:PCR扩增XRCC1启动子序列克隆至pGL3-Basic荧光报告载体上,与E2F1或突变的E2F1(132E)表达载体分别同时转染Sao2细胞;从基因Bank中调取XRCC1启动子序列和E2F结合位点序列分析其相关性;设计引物逐渐从5'端删除与E2F结合位点相关的序列,将PCR产物分别克隆至pGL3-Basic荧光报告载体上,转染至Sao2细胞.细胞裂解后与β-gal反应液一同保温,570 nm处读取吸光度值.结果:XRCC1与E2F1共转染后,可以诱导XRCC1荧光强度的增加;XRCC1启动子序列5'端-819～-803与E2F结合位点相关,删除5'端序列使荧光强度减少.结论:E2F1作用于XRCC1启动子序列,上调XRCC1转录.     

重叠延伸PCR法构建UGRP1基因启动子的点突变表达载体

背景与目的:构建UGRP1基因启动子的定点突变表达载体.材料与方法:以插入UGRP1基因正常启动子的质粒pGL3-UGRP1(-112G)为模板,用重叠延伸PCR定点诱变技术,对-112位点的碱基进行定点突变,并构建定点突变表达载体.结果:DNA测序表明,UGRP1基因启动子-112处的碱基已由G突变为A,成功实现定点诱变.结论:重叠延伸PCR定点诱变技术高效、简便.pGL3-UGRP(-112A)的成功构建,为进一步研究-112G/A多态性对该基因转录活性的影响奠定了基础.     

人类生存蛋白启动子的克隆及其在淋巴瘤细胞中的转录活性研究

  目的　克隆人类生存蛋白（Survivin）核心启动子,研究Survivin启动子在人类淋巴瘤细胞Ramos和健康人肝脏细胞Chang Liver中的转录活性。方法　以人类肠基因组DNA为模板,PCR扩增Survivin启动子987 bp片段,将其通过酶切位点连入pGL3-Basic载体构建pGL3-Survivin荧光素酶报告基因载体,通过脂质体转染法转染Ramos和Chang Liver 细胞,通过检测荧光素酶表达水平,比较Survivin启动子在这两种细胞中的转录活性。结果　成功克隆出987 bp的Survivin启动子;双酶切、PCR检测和DNA测序证实pGL3-Survivin载体构建成功;荧光素酶活性检查显示：Survivin启动子在Ramos细胞中的转录活性为阳性对照CMV启动子活性的4.5 %,明显高于在Chang Liver细胞中的0.19 %,在淋巴瘤细胞中具有较高特异性,且在淋巴瘤细胞中的活性明显高于阴性对照pGL3-Basic的0.086 %。结论　Survivin启动子在淋巴瘤细胞中具有较高的特异性,可以作为淋巴瘤细胞基因转染的肿瘤特异性启动子使用。     

MTS1基因β启动子E2F1结合位点序列突变重组质粒的构建与表达

目的：深入研究MTS1基因β启动子的转录激活与E2F1转录因子的相互作用关系，阐明该转录水平的调控机制。方法：用PCR定点突变方法或酶切连接法，构建β启动子0.38kb SacⅡ-Sac I酶切片段中E2F1 A，B，C任意2个位点或3个位点均突变的pGL3重组质粒。用脂质体介导的基因瞬时转染法，将构建的重组质粒转染MTS1基因双等位缺失的急性T淋巴细胞白血病Jurkat细胞，检测pGL3重组质粒中荧光素酶报告基因的表达。结果：构建的E2F1 A，B，C结合位点突变的重组质粒经Sac I或Nae I酶切鉴定和DNA序列分析得到证实。与E2F1位点野生型重组质粒比较，突变型重组质粒在Jurkat细胞中荧光素酶报告基因的表达量减少，以3个位点均突变的重组质粒为明显。结论：构建的E2F1 A，B，C2个或3个结合位点均突变重组质粒成功，可通过基因转染用于研究MTS1基因的功能试验中；MTS1基因β启动子的转录活性可能与E2F1转录因子的反式激活有关。     

hTERT启动子的克隆及hTERT启动子/SV40增强子在食管癌细胞中的转录活性

目的 克隆hTERT启动子核心序列,研究hTERT启动子/SV40增强子在食管癌细胞中的联合转录活性。 方法 以人基因组DNA为模板,PCR扩增hTERT启动子核心片段;将其分别插入荧光素酶基因报告质粒pGL3-Basic和pGL3-Enhancer中,构建hTERT启动子调控的表达载体pGL3-hTERTp和由hTERT启动子/SV40增强子联合调控的表达载体pGL3-hTERTp-SV40en,将上述重组质粒分别瞬时转染食管癌细胞Eca-109、EC1和人胚肺成纤维细胞MRC-5,用荧光素酶检测试剂盒检测转染细胞中荧光素酶基因的表达水平并以此计算hTERT启动子和hTERT启动子/SV40增强子在各种细胞中的转录活性。结果 克隆出长213 bp的hTETR启动子核心片段,DNA测序结果与GenBank中hTERT启动子的碱基序列完全一致;成功构建真核表达载体pGL3-hTERTp和pGL3-hTERTp-SV40en;hTETR启动子在食管癌细胞Eca-109和EC1中均有转录活性,在MRC-5细胞中无明显转录活性;hTERT启动子/SV40增强子在食管癌细胞Eca-109和EC1中的转录活性显著高于hTETR启动子的单独转录活性。结论 hTERT启动子在食管癌细胞中具有靶向性转录活性,SV40增强子能显著增强hTERT启动子在食管癌细胞中的转录活性,有可能作为肿瘤靶向性基因治疗的转录调控元件。     

人端粒酶催化亚单位(hTERT)启动子的克隆及其在人肺癌细胞中转录活性的研究

目的克隆人端粒酶催化亚单位(hTERT)的启动子，并检测它在多种人肺癌细胞株和人胚肺成纤维细胞株中的转录活性，为肺癌靶向性基因治疗的研究奠定基础。方法以人胚肾293细胞基因组DNA为模板，应用PCR方法克隆hTERT 5’端上游旁侧序列长约1．1kh的启动子片段，经DNA测序无误后克隆人荧光素酶报告质粒pGL3-Basic的荧光素酶基因上游，构建pGL3-hTER Tp重组质粒，用脂质体法瞬时转染人肺癌细胞株A549、SPC-A-1、LTEPa-2、NCI—H446、YTMLC、GLC-82、A2，以及人胚肺成纤维细胞株MRC5，转染48h后检测hTERT启动子在各细胞株中的转录活性。结果琼脂糖凝胶电泳显示PCR克隆的hTERT启动子片段长约1.1kb。DNA测序结果与GenBank中hTERT启动子DNA序列完全一致，其5’端和3’端分别位于hTERT基因转录起始位点上游1126bp和43bp，片段长度为1084bp。采用双酶切和PCR两种方法鉴定pGL3-hTERTp重组质粒，均显示构建成功。瞬时转染及荧光素酶活性检测实验显示，hTERT启动子在所检测的肺癌细胞株中均有高低不同的转录活性，而在MRC-5细胞株中无转录活性。结论该实验克隆的1084bp大小的hTERT启动子在多种肺癌细胞株中均有转录活性，在人胚肺成纤维细胞中无转录活性。hTERT启动子有可能作为调控元件用于肿瘤靶向性基因治疗。     

人脂肪酸合成酶FAS启动子的克隆及其在几种肿瘤细胞中的转录活性分析

目的:克隆人脂肪酸合成酶基因FAS启动子的序列,构建重组荧光素酶表达载体pGL3-FAS,并分析其在几种肿瘤细胞中的转录活性,为其作为肿瘤靶向基因治疗的工具提供依据。方法:以人基因组DNA为模板,PCR扩增FAS启动子区680bp活性序列,经测序鉴定正确后,克隆至荧光素酶表达载体pGL3-enhancer中,构建成重组荧光素酶表达载体pGL3-FAS。将pGL3-FAS通过脂质体转染胃癌细胞系SGC-7901、人宫颈癌细胞系Hela、人乳腺癌细胞系SKBR3、人肝癌细胞系HepG2以及NIH3T3成纤维细胞系中,应用荧光素酶检测系统测定荧光素酶活性,通过内参校正得到相对转录活性。结果:成功扩增出大小约为680bp的FAS启动子序列,经测序鉴定与Genebank报道的一致。经酶切鉴定,成功构建重组荧光素酶表达载体pGL3-FAS。瞬时转染pGL3-FAS,发现其在SGC-7901、Hela、SKBR3、HepG2细胞中均具有较强的荧光素酶活性,且荧光素酶活性高于强启动子SV40驱动的pGL3-control载体;而在正常成纤维细胞中,转录活性较低。结论:FAS启动子在肿瘤细胞中具有强转录活性,而在正常细胞中转录活性很低,具有良好的肿瘤靶向性,为肿瘤靶向基因治疗提供理论依据。     

E2F1对XRCC1启动子调节作用的研究

目的：探讨E2F1对XRCC1启动子的调节作用及其意义。方法：PCR扩增XRCC1启动子序列克隆至pGL3-Basic荧光报告载体上，与E2F1或突变的E2F1（132E）表达载体分别同时转染SaO2细胞；从基因Bank中调取XRCC1启动子序列和E2F结合位点序列分析其相关性；设计引物逐渐从5′端删除与E2F结合位点相关的序列，将PCR产物分别克隆至pGL3-Basic荧光报告载体上，转染至SaO2细胞。细胞裂解后与β-gal反应液一同保温，570nm处读取吸光度值。结果：XRCC1与E2F1共转染后，可以诱导XRCC1荧光强度的增加；XRCC1启动子序列5′端-819～-803与E2F结合住点相关，删除5′端序列使荧光强度减少。结论：E2F1作用于XRCC1启动子序列，上调XRCC1转录。     

hTERT启动子克隆及其肿瘤细胞特异性的研究

目的:探讨克隆人端粒酶逆转录酶(hTERT)启动子对肿瘤细胞特异性的意义.方法:全基因合成hTERT启动子核心序列TP258,克隆入T载体,构建pUCmT-TP258测序;Sal Ⅰ BamH Ⅰ酶切pUCmT-TP258,将其片断插入pGL3-Basic/Xho I Bgl Ⅱ位点,构建pGL3-TP258质粒载体.将pGL3-Basic、pGL3-control和pGL3-TP258转染培养的人结肠癌细胞株(CaCo-2)、人乳腺癌细胞株(MCF-7)、原代肾癌细胞(RCC)和人正常成纤维细胞(UFC、BJ、MRC-5),测定Luciferase活性,计算其相对活性.结果:在端粒酶阳性的肾癌细胞RCC、结肠癌细胞CaCo-2和乳腺癌细胞MCF-7中,TP258启动子相对活性很高,分别为32.40±15.32、37.70±6.38和12.80±7.28;而在正常的UFC、BJ和MRC-5细胞中,TP258活性很低,分别是0.40±0.14、0.26±0.13和0.38±0.05,两组闻差异有统计学意义,P=0.003 5.结论:hTERT核心启动子TP258在肿瘤细胞中活性明显高于在正常细胞中的活性,具有肿瘤特异性.TP258可以介导基因在肿瘤中的定向表达,实现基因表达的靶向性.     

新疆地区宫颈癌和CIN患者HPV-16感染和血清中HPV-16 L1抗体的检测及其临床意义

目的 探讨宫颈癌和宫颈上皮内瘤变（CIN）患者血清中HPV 16 L1抗体水平及其与HPV-16感染的关系。方法 收集宫颈癌患者51例和CIN（包括CINⅠ、CINⅡ、CINⅢ）患者44例。取患者宫颈病变组织并提取组织DNA,PCR检测HPV-16 DNA。同时取患者血样,酶联免疫吸附试验（ELISA）检测血清样品中HPV-16 L1抗体。结果 宫颈癌组中HPV-16 DNA的阳性率为82.4％,CIN组为52.3％。宫颈癌组血清HPV-16 L1抗体阳性率为70.6％,CIN组为79.5％。宫颈癌组中HPV-16 DNA阳性而HPV-16 L1抗体阴性的比例为27.5％,显著高于CIN组的9.1％（P＜0.05）;宫颈癌组HPV-16 DNA阴性而HPV-16L1抗体阳性的比例为15.7％,显著低于CIN组的36.4％（P＜0.05）。宫颈癌组HPV-16 DNA和患者HPV-16 L1抗体均阳性的重合率为54.9％,CIN组为43.2％。结论 CIN和宫颈癌患者HPV-16 L1抗体的产生与HPV-16 DNA的检出率相关,血清中HPV-16 L1抗体可作为宫颈癌和CIN病程的辅助诊断指标。     

Distribution of HPV-16 Intratypic Variants among Women with Cervical Intraepithelial Neoplasia and Invasive Cervical Cancer in Mongolia

Objectives. Infection with high-risk human papillomavirus (HPV) is a critical factor associated withcarcinogenesis of the uterine cervix. HPV-16 is most frequently found, and is further subclassified into intratypicvariants based on the nucleotide sequences of the viral genes. Although certain HPV-16 variants are reported tobe associated with the progression of cervical lesions, these relationships remain controversial with differentresults for different populations. To provide data for another population, we investigated the prevalence ofHPV-16 and distributions of its intratypic variants among Mongolian women with cervical intraepithelialneoplasia (CIN) and invasive cervical cancer. Materials and Methods. We analyzed samples from 374 randomlyselected women who attended the National Cancer Center of Mongolia between January 2002 and July 2007,including 147 invasive cervical cancer patients, 127 CIN patients and 100 age-matched controls who werecytologically normal. HPV genotyping was initially conducted, followed by variant analysis for HPV-16-positivesamples by nucleotide sequencing of the E6 gene. The HPV data were evaluated statistically for correlationswith the patients’ clinical data. Results. HPV genotyping detected 101 HPV-16-positive samples. Among thesesamples, 92 were available for subsequent variant analysis, including 66 invasive cervical cancer samples, 25CIN samples and 1 cytologically normal sample. A total of 14 different variants were identified. All 14 variantsbelonged to the European lineage, and the European prototype was detected in 66% (61/92) of the samples.Among the remaining 31 variants, variants with the T350G nucleotide change were predominant (13/31, 42%),followed by variants containing G94A (11/31, 35%), G176A (4/31, 13%) and G274T (2/31, 7%) . There were nosignificant differences among all the variants regarding their distributions in CIN and invasive cervical cancers.Conclusions. HPV-16 variants of the European lineage were exclusively distributed among the Mongolian womenexamined, and the European prototype was overwhelmingly predominant. Since no significant differences werefound between the types of variants and severities of the cervical lesions, it is possible that racial or geographicfactors may have some influences on these relationships.     

Rs10993994对前列腺癌细胞MSMB基因转录的影响

背景与目的：全基因组相关联研究发现了染色体10q11区域的单核苷酸多态性(single nucleotide polymorphism,SNP)位点rs10993994与前列腺癌的遗传易感性相关,本研究探讨rs10993994影响前列腺癌发病风险的可能机制,即研究rs10993994对前列腺癌细胞微精液蛋白β(microseminoprotein beta,MSMB)基因转录活性的影响。方法：化学合成MSMB基因的启动子序列,由于位于序列中间的SNP位点rs10993994有2种等位基因(T/C)可能,合成2条启动子MSMB promoter-T和MSMB promoter-C。将2条启动子通过酶切的方法导入荧光素酶报告基因载体(pGL3-basic vectors)中,筛选阳性克隆后,将携带启动子的质粒转染入前列腺癌细胞PC-3和LNCaP中,最后通过荧光检测仪检测重组质粒中启动子的转录活性。结果：成功合成2条启动子序列：MSMB promoter-T和MSMB promoter-C;分别将启动子序列与荧光素酶报告基因载体重组,形成重组质粒pGL3-MSMB promoter-T和pGL3-MSMB promoter-C。在前列腺癌细胞PC-3中,重组质粒MSMB promoter-C组的相对荧光度为2.27±0.39,显著高于重组质粒MSMB promoter-T组(0.57±0.13),差异有统计学意义(P＜0.05);在前列腺癌细胞LNCaP中,重组质粒MSMB promoter-C组的相对荧光度为1.70±0.32,显著高于重组质粒MSMB promoter-T组(0.37±0.09),差异有统计学意义(P＜0.05)。结论：重组质粒pGL3-MSMBpromoter-C的转录活性强于重组质粒pGL3-MSMB promoter-T的转录活性;rs10993994可以影响到MSMB基因启动子的启动转录活性,rs10993994位点上等位基因胞嘧啶(C)较胸腺嘧啶(T)更能促使MSMB基因转录。     

Enhancement of human papilloma virus type 16 E7 specific T cell responses by local invasive procedures in patients with (pre)malignant cervical neoplasia

It has been suggested that local invasive procedures may alter the natural course of (pre)malignant cervical disease. This could be due to partial excision of the lesions, or via induction of cellular immunity against human papillomavirus (HPV) by the local invasive procedures. We studied the influence of local invasive procedures on HPV-16 E7 specific immune responses in patients with different grades of cervical intra-epithelial neoplasia (CIN) and different stages of cervical cancer. Blood was obtained at intake and after invasive procedures from patients with CIN or cervical cancer. Antigen specific T-cell responses were measured by IFN-gamma ELISPOT analysis, after stimulation with recombinant HPV-16 E7 protein. As expected, HPV-16 E7 specific IFN-gamma T cell responses were more frequent in HPV-16 DNA positive patients compared with that in HPV-16 DNA negative patients (39/50 vs. 16/36, (p=0.006, chi2 test). After invasive procedures, a small number of HPV-16 DNA positive CIN patients, but a considerable proportion of HPV-16 DNA positive cervical cancer patients, showed an enhancement of T cell responses against HPV-16 E7. Induction of T cell reactivity was most pronounced in cervical cancer patients who had undergone previous invasive procedures. Both CD4+ and CD8+ T cells showed E7 specific IFN-gamma production upon in-vitro stimulation. Our study shows that invasive procedures may enhance HPV-specific cell-mediated immunity in a considerable number of patients with cervical cancer, but in only a minority of CIN patients. Our data indicate that invasive procedures should be considered as possible confounding factors when analyzing the effectiveness of therapeutic immunization studies, especially, when induction of HPV-specific immune responses is used as intermediate end-point.     

新疆维吾尔族妇女宫颈癌HPV感染型别分布研究

目的探讨人乳头瘤状病毒(HPV)在新疆南部维吾尔族妇女宫颈癌患者的型别分布情况,为开发适宜该地区的HPV疫苗提供一定的理论依据.方法收集2008年6月至2010年4月就诊于新疆维吾尔自治区人民医院妇科的经病理确诊的新疆南部地区维吾尔族妇女宫颈癌患者120例,利用聚合酶链反应(PCR)和基因芯片技术检测HPV DNA并分...     

多重实时PCR在宫颈癌及癌前病变HPV-16病毒存在状态检测中的应用

Objective: To explore an ideal approach for detecting the physical status of HPV-16 in clinic use and to investigate the integrated HPV-16 in CINs and cervical cancer. Methods: Multiplex real-time PCR method was established to quantify the copy numbers of E2 and E6 genes (E2/E6) for analysis of the physical status of H PV-16 DNA and this assay was compared to Southern blot analysis. HPV-16-containing paraffin-embedded tissues including 49 CINs and 51 cervical squamous cancers were detected using the method. Results: (1) The cutoff ratio of E2/E6 to distinguish pure episomal from mixed H PV-16, was0.81 in the multiplex real-time PCR; (2) The agreement rate between multiplex real-time PCR and Southern blot was 81.5%(the Kappa statistic was 0.844, P＜0.001); (3) HPV-16 DNA existed in an episomal form in 57.1% and mixed form in 42.9%of CIN Ⅰ lesions; The concomitant form of HPV-16 (＞70%) constituted the majority in CIN Ⅱ and CIN Ⅲ; HPV-16 DNA mostly integrated into the host chromosome (s) in squamous cervical cancers (68.6%); (4) The incidence of HPV-16 integration was increased with the degree of cervical lesions; (5) The frequency of pure integrated HPV-16 in stage Ⅱ Ⅲ (88%) was significantly higher than that in stage Ⅰ (33.3%). Conclusion: (1) Mutiplex real-time PCR provides a rapid, sensitive and reliable method for clinic detection of the physical state of HPV-16 DNA; (2) The integration of the HPV-16 DNA is a very early and important event in the progression from preinvasive to invasive cervical cancer; (3) The pure integrated status of HPV-16 in cervical cancer may be associated with poor prognosis of cervical cancer, but further study will be needed to prove its prognostic significance.     

人乳头瘤病毒16型转化基因在不同阶段宫颈上皮病变组织中的分布及基因变异特点

目的 研究人乳头瘤病毒(HPV)16 E6、E7和E5基因在湖北地区不同阶段宫颈上皮病变患者组织中的分布以及E6、E7基闪的变异特点.方法 从124例宫颈癌、17例宫颈上皮内瘤变(CIN) Ⅰ+CINⅡ级、23例CIN Ⅲ级和36例慢性宫颈炎患者活检或手术切除标本中提取组织DNA,用HPV16 E6、E7和E5特异性引物进行PCR扩增,对部分扩增的 E6 和 E7 产物片段进行测序分析.结果 在官颈炎、CIN Ⅰ+CINⅡ级、CINⅢ级和宫颈癌组织中,E6基因的阳性率分别为25.0%、29.4%、60.9%和76.6%;E7基因的阳性率分别为16.7%、41.2%、43.5%和61.3%:E5 基因的阳性率分别为5.6%、5.9%、30.4%和40.3%.E6、E7和E5基因在不同阶段宫颈上皮病变组织中的阳性率差异均有统计学意义(均P<0.01).在80例官颈癌测序组织中,有47例发生E6基因178位点的T→C突变,突变率为58.8%,相应氨基酸由天冬氨酸(Asp)改变为谷氨酸(Glu);而在20例宫颈炎和22例CINⅠ～Ⅲ级测序组织中,E6基因178位点的突变率分别为25.0%和31.8%.在30例宫颈癌测序组织中,有21例发生E7基因647位点的A-G突变,突变率为70.0%,相应氨基酸由天冬酰胺(Asn)改变为丝氨酸(Ser);而在20例宫颈炎和22例CINⅠ～Ⅲ级测序组织中,E7基因647位点的突变率分别为35.0%和40.9%.结论 HPV16 E6、E7和E5基因与宫颈癌的发生和发展有高度的相关性.但E5基因在不同阶段官颈上皮病变中可能存在不同程度的缺失.中国湖北地区流行的HPV16病毒株可能为HPV16亚洲型变异株.     

Decreased expression of human papillomavirus E2 protein and transforming growth factor-beta1 in human cervical neoplasia as an early marker in carcinogenesis

BACKGROUND AND OBJECTIVES: Human papillomavirus (HPV) is thought to be one of the possible causative factors in cervical carcinogenesis, and cervical carcinoma cells are refractory to tumor transforming growth factor (TGF)-beta1. The purpose of this study is to investigate the possible cause-effect association between HPV and TGF-beta1 during cervical tumorigenesis. METHODS: We assessed the expression of HPV capsid proteins, HPV-16 E7, HPV-16 E2 (C and N terminals), TGF-beta1, and their receptors TGF-beta RI and RII by immunohistochemistry in 48 paraffin-embedded blocks of tumor tissue derived from patients of cervical neoplasia. RESULTS: Expression of TGF-beta1 decreased as tumor cells progressed from cervical intraepithelial neoplasia (CIN)1, CIN2, CIN3, to microinvasive carcinoma (P < 0.05). Levels of TGF-betaRI and TGFbeta-RII stayed the same in all cases. HPV was found in 89.6% of the studied sections, and cervical lesions without HPV infection expressed significantly less TGF-beta1 (P < 0.05). By comparing the expression pattern of TGF-beta1 and HPV in the neoplastic cells with that of normal cervical epithelium in each section, we found loss of HPV-16 E2 higher in CIN3 (15/24) than in CIN1 or CIN2 (3/7), and there is a significant trend that loss of HPV-16 E2 expression correlated with a >50% loss of TGF-beta1 at the lesion site (P < 0.05). CONCLUSIONS: Our result showed co-suppression of HPV and TGF-beta1 expression during progression of cervical squamous cell cancer. Using antibody against HPV-16 E2 may be an auxiliary tool for the investigation of cervical tumor progression.