Antibodies to Herpes simplex virus types 1 and 2 in patients with squamous-cell carcinoma of uterine cervix in India.

Antibody activity to Herpes simplex virus type-1 (HSV-1) and type-2 (HSV-2) was measured by the indirect hemagglutination (IHA) test in sera from 124 women with squamous-cell carcinoma of the uterine cervix, 46 women with non-cervical cancer and 116 matched normal women. The type-specific antibodies were assayed by IHA inhibition test. The proportion of cervical cancer cases positive for HSV-2 infection was significantly greater than that of non-cervical cancer cases and matched control women. No difference was observed in the percentages positive for HSV-1 infection among the three study groups. The mean titers of antibodies to HSV-1 as well as HSV-2 were significantly higher than the respective titers in the other two study groups. However, determination of type-specific antibodies did not help in establishing evidence for past infection by HSV-2 in patients with cervical cancer, as no type-specific HSV-2 antibodies were produced in 58% of the cases. The cervical cancer cases were also analysed according to clinical staging (Stages 0 to IV) and histological differentiation of squamous-cell carcinoma. Neither the percentage of positives for HSV-2 infection nor HSV-antibody titers varied, either in clinical staging or in histological differentiation of cervical cancer. These findings suggest that HSV-2 may be related to cervical cancer as an etiological agent but not as a passenger virus of carcinomatous tissue.     

宫颈癌病毒病因中人乳头病毒和疮疹病毒：II型相关性的研究

HPV DNA and HSV-2 antigens were detected in a series of cervical lesions by DNA hybridization and PAP stain techniques in order to investigate the possible role of HPV and HSV-2 in the viral etiology of cervical carcinoma. The correlation of HPV DNA sequence and HSV-2 antigens was analyzed. The results showed that the positive rates of HPV 16, 18 and 11 in cervicitis, dysplasia and cervical cancer were 13%; 0, 13%; 17%, 17%, 17%; 43%, 8%, 4%; and HSV-2 antigens were found in 41% of cervicitis, 50% of cervical dysplasia and 63% of cervical carcinoma. Comparing the results of HPV hybridization with that of HSV-2 antigens, the authors found that both HPV DNA and HSV-2 antigens could be detected in some cases. The results indicate that the viral etiology of cervical carcinoma may be multifactorial and HSV-2 and HPV may be synergistic in the development of cervical carcinoma.     

Herpes simplex virus-2 as a human papillomavirus cofactor in the etiology of invasive cervical cancer

BACKGROUND: Human papillomavirus (HPV) infection is the main cause of invasive cervical cancer, but cofactors may act in conjunction with HPV. We performed a pooled analysis of seven case-control studies to examine the effect of one possible HPV cofactor, herpes simplex virus-2 (HSV-2) infection, in the etiology of invasive cervical cancer. METHODS: Blood and exfoliated cervical specimens were obtained from 1263 case patients with invasive cervical cancer (1158 with squamous-cell carcinomas and 105 with adeno- or adenosquamous-cell carcinomas) and 1117 age-matched control subjects. Western blot analysis and/or an enzyme-linked immunosorbent assay were used to detect type-specific serum antibodies to HSV-2 and HSV-1, and Chlamydia trachomatis serum antibodies were detected using a micro-immunofluorescence assay. HPV DNA was detected using a polymerase chain reaction assay. Summary odds ratios (ORs) and 95% confidence intervals (CIs) were computed from unconditional logistic regression models. RESULTS: Overall, HSV-2 seropositivity was higher among case patients with squamous-cell carcinoma (44.4%, 95% CI = 41.5% to 47.3%) or adeno- or adenosquamous-cell carcinoma (43.8%, 95% CI = 34.2% to 53.5%) than among control subjects (25.6%, 95% CI = 23.0% to 28.2%). Cervical specimens from 1098 (94.8%) squamous-cell carcinoma case patients, 95 (90.5%) adeno- or adenosquamous carcinoma case patients, and 164 (14.7%) control subjects were positive for HPV DNA. Among the HPV DNA-positive women, HSV-2 seropositivity was associated with increased risks of squamous-cell carcinoma (OR = 2.19, 95% CI = 1.41 to 3.40) and adeno- or adenosquamous-cell carcinoma (OR = 3.37, 95% CI = 1.47 to 7.74) after adjustment for potential confounders. A similar association between HSV-2 seropositivity and squamous-cell carcinoma risk was observed after further controlling for markers of sexual behavior (OR = 1.96, 95% CI = 1.24 to 3.09). Among control subjects, HSV-2 seropositivity was associated with markers of sexual behavior, but not with cervical HPV DNA positivity. CONCLUSION: HSV-2 infection may act in conjunction with HPV infection to increase the risk of invasive cervical carcinoma.     

Herpes simplex virus type 2 and human papillomavirus type 16 in cervicitis, dysplasia and invasive cervical carcinoma

Sera and biopsies of cervical lesions from 55 women with benign or malignant disease were analyzed for evidence of infection with herpes simplex virus type 2 (HSV-2) or human papillomavirus (HPV). In addition, information regarding known risk factors for cervical cancer was obtained by interview. The sera were tested for HSV-2 antibodies and the biopsies were tested for HPV or HSV DNA sequences by Southern blot hybridization. HSV-2 sequences were detected in 2/13 (15%) invasive neoplasms and in 1/12 (7%) benign lesions. Under non-stringent conditions of hybridization, reactions with HPV DNA were detected in biopsies of 2/17 (12%) inflammatory lesions, 6/12 (50%) intraepithelial neoplasms and 13/20 (65%) invasive neoplasms. All but one of the positive biopsies of invasive cancer, but only 4/11 biopsies of non-invasive lesions, contained HPV-16 DNA as determined by stringent hybridization conditions. Women with cervical cancer possessed the risk factors associated with the disease. Cigarette smoking and the presence of HPV-16 DNA were the most prominent risk factors. No evidence of an interaction between HSV-2 and HPV-16 was found among the cases of invasive cervical cancer.     

Simultaneous presence of herpes simplex and human papilloma virus sequences in human genital tumors

Herpes simplex virus (HSV) and human papillomavirus (HPV) sequences were analyzed in tumors of the female lower genital tract, by probing DNA from 13 intraepithelial and 30 invasive neoplastic lesions with radiolabelled HPV-16 and HPV-18 DNA as well as cloned fragments of HSV-2 DNA. Careful removal of stromal tissue from the pathological specimens allowed authentic tumor DNA to be processed. Normal genital tissue obtained from the patients and genital condylomata were included as internal controls. The presence of HPV-16 or 18 DNA was detected in 12/13 (92.3%) intraepithelial neoplasms and in 16/30 (53.3%) invasive carcinomas. No significant difference was detected in titer or frequency of antibodies to HPV group-specific antigen in sera from patients and controls. Hybridization to BgIII N fragment of HSV-2 DNA was detected in 4/13 (30.8%) intraepithelial neoplasms and 4/30 (13.3%) invasive carcinomas but in none of the control tissues. All the 8 samples harboring HSV-2 homologous sequences were also positive for HPV, supporting the hypothesis of a synergistic association between the 2 viruses. The hybridization analyses performed to study c-myc involvement in genital oncogenesis did not reveal c-myc amplification in either invasive or pre-invasive lesions.     

宫颈癌中HPV16 E5基因的变异与病毒物理状态的研究

目的研究HPV16DNA在宫颈癌组织中的物理状态及其与E5转化基因的变异的关系。方法用斑点杂交方法确定HPV16阳性的宫颈癌标本,采用Southern印记杂交技术检测HPV16DNA在宫颈癌组织中的物理状态,并结合E5基因的扩增及序列测定来综合探求HPV16基因组在宿主细胞中的物理状态及其与E5片段变异的关系。结果60例宫颈癌组织中HPV16阳性率为58%(35/60);其中有22%(8/35)的HPV16DNA是以游离状态存在于宿主细胞内。在HPV16以游离型存在的宫颈癌标本中,仅有1例(1/8)发生了变异。结论不同病理分期的宫颈癌组织HPV16的物理状态无明显差异。在游离型HPV16致癌机制中,E5的变异似乎不占主导地位。所得结果为揭示HPV16致宫颈癌的分子机制尤其是为E5在游离型HPV16致癌机制提供了新资料。     

Serum antibodies to herpesvirus early antigens in patients with cervical carcinoma determined by anticomplement immunofluorescence technique.

Sera from patients with carcinoma of the uterine cervix were used with the anticomplement immunofluorescence (ACIF) technique to develop a simple procedure for detection of antibodies to herpes simplex virus type 2 (HSV-2) early antigens. Test cells used in the ACIF assay were HEp-2 cells infected with HSV-2 sequentially treated with inhibitors of protein and of DNA SYNTHESIS. The cells were first treated for 7 h with cycloheximide (100 mug/ml) and then for 3 h with hydroxyurea (150 mug/ml). In this assay serum titers giving more than 21.5% ACIF-positive cells correlated to invasive carcinoma of the cervix. Using these criteria 18 out of 43 patients with invasive carcinoma, none of 8 patients with carcinoma in situ, and only 1 of 43 controls, were diagnosed as positive. Thus an elevated titer of antibodies against HSV-2 antigens correlated to cervical carcinoma.     

HPV 16 in multiple neoplastic lesions in women with CIN III

Paraffin embedded material of multiple primary cancers and other hyperplastic tumours from fifteen patients were analyzed by PCR and in situ hybridization for the presence of HPV DNA in the lesions. All patients had also high grade cervical intraepithelial dysplasia (CIN III) and breast carcinomas and were selected from a previous study enrolling 46 women with CIN III and breast carcinomas. HPV 16 was detected by PCR in 8/15 patients (53%), with eleven HPV 16 positive tumours. HPV 16 was detected in two malignant melanomas, one basal cell carcinoma, one squamous cell carcinoma of the vulva, one Bowen disease of the vulva, two high grade vaginal intraepithelial neoplasias, one cancer corporis uteri, one bronchial carcinoma and two lymphomas. Three cases, two high grade vaginal intraepithelial neoplasia and a squamous cell carcinoma of the vulva, were also reported to be positive by in situ hybridization. 5/8 patients (63%) with HPV 16 positive second cancers had also HPV 16 positive breast carcinomas. All fifteen patients with second cancers after CIN III had HPV 16 positive CIN III lesions; 53% of the patients had also a familial cancer history. We assume that HPV 16 may be involved in the development of different second cancers in women with HPV 16 positive CIN III.     

THE CERVIX MULTI－VIRUSES INFECTION AND THE DEVELOPMENT OF CERVICAL CARCINOMAS

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Herpes simplex virus type 2: a possible interaction with human papillomavirus types 16/18 in the development of invasive cervical cancer.

A case-control study of 766 histologically confirmed incident cases of invasive cervical cancer and 1,532 hospital and community controls was conducted in Latin America to evaluate the etiologic role of herpes simplex virus type 2 (HSV-2) and to examine whether HSV-2 interacts with other risk factors. In addition to a personal interview, all subjects were asked to donate blood samples and cervical swabs for assessment of exposure to HSV-2 and human papillomaviruses (HPVs) respectively. Ninety-eight percent of cases and 91% of controls agreed to the interview and blood collection. Women testing positive for HSV-2 antibodies were found to have a 60% increased risk of cervical cancer compared with seronegative women (95% CI = 1.3, 1.9). Control for education, sexual and reproductive behavior, prior Pap-smear screening, smoking, oral contraceptive use, HPV-6/11 DNA, or HPV-16/18 DNA detection did not materially affect this estimate. No effect modification of HSV-2 by age, HPV-6/11 DNA, pregnancies, oral contraceptive use or cigarette smoking was observed. However, a significant interaction was detected between HSV-2 and HPV-16/18. Compared with women testing negative to both virus types, those positive for HSV-2 alone had a RR of 1.2 (95% CI = 0.9, 1.6), those positive for HPV-16/18 DNA alone had a RR of 4.3 (95% CI = 3.0, 6.0), and those positive for both viruses had a RR of 8.8 (95% CI = 5.9, 13.0). These findings corroborate recent laboratory evidence of a possible biological interaction between HSV-2 and HPV-16/18 in the development of cervical cancer. Further confirmatory studies are needed, given concerns with potential misclassification of exposure by the laboratory assays utilized.     

Detection of hepatitis-B virus-DNA at high-frequency in liver neoplasias using a PCR technique

Hepatocellular carcinoma is the most frequent liver cancer and hepatitis B virus is included among the risk factors for the development of this type of neoplasia. Direct detection of this virus is difficult due to the lack of a simple tissue culture system for growing the virus. Amplification of HBV nucleic acid sequences with the Polymerase Chain Reaction technique leads to the direct detection of the virus, but involves several critical steps and it is prone to false positive results due to inter sample contaminations. We overcame these shortcomings by using a simple boiling method for extracting DNA from histological slides of tissues coupled with double ('nested') PCR amplification. In this study we evaluated the frequency of presence of HBV nucleic acid sequences in samples of neoplastic liver tissues from patients in Greece. We studied 20 DNA samples from hepatocellular carcinomas and we found 11 positive for HBV DNA and 8 DNA samples from hepatoblastomas and we found 3 positive for the viral DNA.     

宫颈癌组织中HPV_(16)DNA基因同源序列的检测

206例不同病理诊断的宫颈脱落细胞和宫颈组织DNA,同探针杂交,检测HPV_(16)DNA同源性序列。结果表明:HPV_(16)DNA探针同宫颈癌脱落细胞杂交(65.2％)和同宫颈癌组织DNA杂交(66.7％),其杂交率无显著差别(P>0.05)。宫颈炎脱落细胞DNA和宫颈炎组织DNA与HPV_(16)DNA探针杂交率间的差异也无统计学意义。然而,宫颈癌和宫颈炎之间无论是脱落细胞或是组织DNA同探针杂交有显著差异(前者为65.2—66.7％,后者为26.1—28.5％)P<0.01。宫颈癌和宫颈炎DNA经酶切电泳后,Southern blot杂交结果提示HPV_(16)DNA整合在癌组织基因组中。     

Detection of human papillomavirus (HPV) DNA in carcinoma of uterine cervix and genital tract diseases in Jiangsu province, China]

Pst-1 cleaved DNA restriction fragment length polymorphism (RFLP) of biopsy samples from 41 patients with squamous cell carcinoma of uterine cervix collected in Jiangsu province, China were examined for HPVs by Southern blot hybridization using HPV 16 DNA as a probe. 20 of the 41 samples were positive for HPVs when hybridized under non-stringent condition. HPV was not detectable in samples collected in the same time period from patients with cervical adenocarcinoma (N = 2), vaginal carcinoma (N = 3), vulval carcinoma (N = 1) and benign cervicitis (N = 8). Of the 20 positive samples, 7 (17.1%) had CHPV X1, a new type of NPV previously discovered is China, 6 (14.6%) had HPV 16, 4 (9.9%) had HPV 31, and in 3 (7.2%) the HPV type is as yet undetermined. Our data indicate that HPV 16 and CHPV X1 may be more closely related to cancer of the uterine cervix in Jiangsu province.     

Lack of association between herpes simplex virus type 2 infection and cervical cancer--Taq Man realtime PCR assay findings

Background: About one third of the human population suffer cancer during their lifetime and morethan 20% of total morbidity is related to neoplasia. Cervical cancer is generally the most common cancerin developing countries and the second most common in women globally. The role of human papillomaviruses viruses in its induction is clear. However, the involvement of hepres simplex virus type 2 (HSV-2) iscontroversial. Therefore a survey was conducted of the prevalence of HSV-2 in patients with cervical cancerand also healthy people with sensitive and quantitative Taq Man real-time PCR assay. Materials and methods:Seventy six formaldehyde fixed paraffin embedded tissue specimens from patients with histologicallyproven history of cervical cancer as well as 150 control blocks were sectioned for deparaffinization andDNA extraction. Results: There was no HSV-2 DNA in our patient specimens but four control samples werepositive, all with a history of hysterectomy. Conclusion: Considering the absence of any positive viral HSV-2DNA in our patients and also the presence of four positive specimens among our controls, we did not findany relationship between the presence of HSV-2 DNA and cervical cancer.     

Relationship between EB virus and nasopharyngeal carcinoma. I. Detection of EBV DNA-related sequences in biopsy specimens of nasopharyngeal carcinoma

Assay of biopsy materials of nasopharyngeal carcinoma (NPC) by nucleic hybridization (dot blot) technique with 32P labelled EBVDNA BamHIW fragment probe (specific radioactivity: 1-3 x 10(7) cpm/micrograms DNA) are described. 57 biopsies of different pathologic types were taken from NPC patients from Beijing and Qing Dao, Shandong province, and 54 control biopsies were taken from patients with other tumors and inflammatory tissues from the nasopharynx and head-neck regions. It was shown that EBVDNA positive reaction was found in 52 out of 57 NPC biopsies (91%), while only 10 positive reaction were observed in 54 control samples (18.5%). There was a significant statistical difference therein (P less than 0.001), but no significant difference of EBV DNA positive rates was found among the different pathological types (P greater than 0.05). Also no significant difference was found in biopsies taken from various geographical regions in both groups.     

Human cytomegalovirus and herpes simplex type 2 virus in normal and adenocarcinomatous prostate glands

Normal prostate, benign prostate hypertrophy (BHP), and prostate adenocarcinoma (ACP) biopsy specimens were analyzed for the presence of human cytomegalovirus (HCMV) and herpes simplex virus type 2 (HSV-2)-specific macromolecules. HCMV DNA homologous sequences were detected in 2 of 13 normal prostate, 2 of 9 BHP, and 3 of 10 ACP tissues, and HSV-2 DNA homologous sequences were found in 1 normal prostate tissue and 2 ACP tissues. In situ DNA-RNA hybridizations indicated HCMV-specific RNA in 3 of 8 BHP and 4 of 9 ACP tissues. No positive in situ hybridization for HCMV RNA was observed in normal prostate tissues. Parallel in situ DNA-RNA hybridization localized HSV-2-specific RNA in only 1 of 8 tumor sections. No HSV-2-specific RNA was observed in sections of normal and BHP tissues. Anticomplement immunofluorescence (ACIF) tests of BHP and ACP specimens showed specific HCMV immunofluorescence in 3 of 8 BHP and 4 of 9 ACP tissues. ACIF results correlate closely with in situ hybridization results and imply some degree of HCMV association with prostatic abnormality. The results also suggest that latent HCMV may be harbored by the human prostate gland.     

Involvement of viral and chemical factors with oral cancer in Taiwan

BACKGROUND: The association between oral squamous cell carcinoma (OSCC) and viral and chemical factors is uncertain. Therefore the correlation of viral and chemical factors with oral cancer in Taiwan was investigated. METHODS: Thirty-seven paraffin-embedded oral cancer biopsies and 36 normal oral tissue specimens were examined by the polymerase chain reaction method for six viruses: HPV, CMV, EBV, HSV-1, HSV-2 and HHV-8. To elucidate the role of arecoline in the oncogenesis of oral cancer, human buccal fibroblasts, oral submucosal fibroblasts and three cancer cell lines KB, GNM and TSCCa were used for MTT cytotoxity assay and flow cytometry DNA content analysis. RESULTS: Two (5.4%) HSV-1-positive and four (10.8%) HPV-positive cases were recognized in oral cancer biopsies. Among the four HPV-positive tissues, two were further typed as HPV-16, one was identified as HPV-18- and HSV-1-positive; and one contained both HPV-16 and HPV-18. One sample presented HSV-1 only. Arecoline, at a concentration lower than 0.8 micro g/ml, increased cell growth (all cell types); at higher concentrations (25-400 micro g/ml) it was cytotoxic. The cell cycle was demonstrated to be altered either by low or high concentrations of arecoline treatment, depending on the cells treated. CONCLUSIONS: The data demonstrated that HPV, HSV-1 and betel quid chewing were significantly associated with OSCC, but HSV-2, CMV, EBV and HHV-8 were not. We suggest that the most determinative factor for oral cancer may be chemical in nature rather than viral infection.     

Search for human papillomavirus, herpes simplex virus and c-myc oncogene in human genital tumors

We tested grade III cervical intra-epithelial neoplasms (CIN III), genital invasive carcinomas and healthy genital tissues for the presence of a number of viral and cellular parameters, considered to be risk factors in genital oncogenesis. The results show that: (I) about 30% of genital tumors had homology to HSV-2 BglII N and/or BamHI E DNA fragments; (2) positivity to HSV-2-specific protein ICP10, encoded by sequences within the HSV-2 BamHI E DNA fragment, was detected in 29/55 neoplastic genital tissues but not in healthy genital tissues or tumors at other sites; (3) HPV-16 DNA was found in about 50% of tumor samples, but none of the positive tissues reacted to HPV capsid protein or to the protein encoded by the HPV-16 E6 ORF; (4) 6/8 tumor samples showing homology to HSV-2 DNA fragments also hybridized to HPV-16 DNA; (5) c-myc amplification was not detected in any of the analyzed samples, but tissues from 4 patients were positive for c-myc expression. None of the considered factors was present in all the analyzed samples; nevertheless, some tissues showed the simultaneous involvement of several parameters, suggesting that a number of risk factors may be involved in different steps of human genital oncogenesis.     

AG-4 complement-fixing antibodies in cervical cancer and herpes-infected patients using local herpes simplex virus type 2

The incidence of anti-AG-4 complement-fixing antibodies in Australian cervical carcinoma (CaCx), herpes simplex virus (HSV) type 1 and 2, and control patients, was investigated using local HSV strains. The local HSV strains (both HSV-IMI and HSV-2MI) were found, by neutralization experiments, to vary from the American prototype strains. All HSV-2-strains tested were able to induce AG-4 in 4-h infected HEp-2 cells. Anti-AG-4-complement-fixing antibodies were detected in 40% of dysplasia patients, 60% of carcinoma-in-situ patients, 75% of CaCx patients, 65% of CaCx post-operative patients, 88% of HSV-2 patients with active lesions, 10% of HSV-1 patients with active lesions, 10--20% of normal patients and 20% of patients with cancer, other than CaCx. The AG-4 test is tumour-specific in that it distinguishes CaCx from other cancer patients tested, but it cannot distinguish HSV-2 patients from CaCx patients.     

染色体2p22的过度扩增及hmsh2基因的高表达与卵巢癌紫杉醇耐药相关性的研究

目的研究染色体特定区域DNA拷贝改变、基因差异表达及其与紫杉醇耐药的关系。方法运用比较基因组杂交(comparativegenomichybridization,CGH)及RTPCR技术,分析卵巢癌紫杉醇耐药株染色体基因组变化和hmsh2基因在卵巢癌组织中的表达。结果CGH结果显示最有意义的变化是卵巢癌紫杉醇耐药株OC3/Tax300细胞染色体2p22过度扩增。RTPCR检测发现hmsh2基因在OC3/Tax300细胞高度表达。hmsh2基因表达阳性率在组1和组2分别为93.9%(31/33)及47.6%(10/21),差异有显著性;在低分化癌为93.3%,显著高于中、高分化癌的54.2%。结论2p22拷贝数的过度扩增及hmsh2基因的高度表达可能与卵巢癌紫杉醇耐药相关。