Type-specific human papillomavirus E6/E7 mRNA detection by real-time PCR improves identification of cervical neoplasia.

DNA-based human papillomavirus (HPV) assays show high sensitivity but poor specificity in detecting high-grade cervical lesions. Assays detecting mRNA of the oncoproteins E6 and E7 show higher specificity but lack either detection of all high-risk HPV genotypes or the capacity to specify the detected genotypes. Therefore, a real-time PCR assay detecting type-specific E6/E7 mRNA was developed and the clinical performance evaluated. A total of 210 cervical LBC (liquid-based cytology) samples from 204 women were analyzed for HPV DNA and mRNA with the in-house real-time PCR as well as PreTect HPV-Proofer. The sensitivity of real-time PCR mRNA detection to identify histologically confirmed CIN2+ (cervical intraepithelial neoplasia, grade 2 or higher) was 0.91, compared to 0.95 for DNA analysis. The specificity was 0.68 compared to 0.38, and the positive predictive value (PPV) was higher for mRNA (0.67 versus 0.52) without any loss in negative predictive value (NPV). The sensitivity of the real-time PCR mRNA test was somewhat higher than that for PreTect HPV-Proofer (0.83 versus 0.75) in analyses for the same genotypes. The specificities were similar (0.76 versus 0.77). In analyses for mRNA of the eight most common genotypes in cervical cancer (HPV16, -18, -31, -33, -35, -45, -52, and -58), the sensitivity of detection of CIN2+ lesions was 0.87 and the specificity 0.74, with a PPV of 0.70. In conclusion, real-time PCR for detection of HPV E6/E7 mRNA transcripts can be a sensitive and specific tool in screening and investigation of cervical neoplasia. The composition of HPV types in mRNA testing needs to be further investigated to optimize sensitivity and specificity.

A population-based 5-year cohort study including all cases of epithelial ovarian cancer in western Sweden: 10-year survival and prognostic factors.

Epithelial ovarian cancer (EOC) is the major gynecologic cancer mortality cause in Sweden. The aim of the present study was to investigate the long-term survival and prognostic factors of a complete population-based 5-year cohort of 682 patients with invasive EOC in western Sweden (population around 1.6 million). Data relating to residual tumor after surgery, FIGO stage, grade, histopathologic subtype, ploidy status, adjuvant chemotherapy (the prepaclitaxel period), and disease state (recurrence and death) were reported to a quality register in a prospectively kept database and were controlled against the Swedish National Cancer Registry for completeness. The median follow-up durations for the prospectively collected data in the Cox analysis and for the survival analysis that was made for all patients were 81 months (range, 52-109 months) and 11.7 years (range, 8.7-14.1 years), respectively. No patient was lost to follow-up. The relative 10-year survival rate was 38.4% (95% confidence interval, 34.5%-42.8%). The median relative survival time was 4.3 years (95% confidence interval, 3.6%-5.2%). In the univariate Cox regression analysis, prognostic significances for age, stage, residual tumor, histopathologic subtype of serous cystadenocarcinoma, grade, CA-125, and ploidy status were seen. In the multivariate analysis, age, stage, residual tumor after surgery, and postoperative CA-125 were of prognostic significance. In conclusion, 4 major prognostic factors were found for EOC in this population-based cohort study that also presents nearly accurate long-term survival owing to the nonselective nature and completeness regarding patients and follow-up of the study.

Effect of adjuvant paclitaxel and carboplatin for advanced stage epithelial ovarian cancer: a population-based cohort study of all patients in western Sweden with long-term follow-up.

OBJECTIVE: To evaluate long-term survival and prognostic factors for all epithelial ovarian cancer (EOC) patients after adjuvant treatment with paclitaxel and carboplatin. DESIGN: Prospectively collected data from a population-based cohort. SETTING: Western Sweden Health Care Region. POPULATION: All women diagnosed with EOC between 1998 and 2005. METHODS: Data related to age, stage, surgery, histopathology, grade, ploidy status, CA-125, follow-up, recurrence and death of EOC patients (n=976) were prospectively collected in a quality register. No patient was lost to follow-up and the median follow-up was 68 months (range: 27-110). MAIN OUTCOME MEASURES: Relative survival at 5 and 8 years for all and for those treated with chemotherapy; median progression-free survival (PFS) for stage IIB-IV patients treated with paclitaxel and carboplatin. RESULTS: Relative 5- and 8-year survival rates in the subgroup of patients treated with chemotherapy after surgery (n=853) were 50.4% (95% CI: 46.4-54.3) and 40.5% (95% CI: 35.4-45.6), respectively. The median relative survival time of the entire group of patients was 60 months (95% CI: 52-73). The median PFS for the patients in stage IIB-IV treated with paclitaxel and carboplatin was 18 months (95% CI: 17-20). Well-established prognostic factors of age, stage, residual tumor and post-operative CA-125 were of prognostic significance. CONCLUSION: Post-surgical adjuvant chemotherapy of paclitaxel and carboplatin for advanced stages of EOC does not seem to increase the relative 5-year survival rate or the median PFS compared to results of earlier studies of a similar patient cohort from the same geographical area.

Genetic alterations of serous borderline tumors of the ovary compared to stage I serous ovarian carcinomas.

Borderline tumors of the ovary comprise 10-20% of all epithelial ovarian tumors, and are placed between clearly benign and obviously malignant ovarian tumors. The issue of whether borderline tumors are precursors of invasive carcinoma or distinct clinical entities, however, is still the subject of discussion. To increase our understanding in relation to this issue, the aim of our study was to analyze both serous borderline and invasive ovarian tumors, and to investigate early carcinogenesis in serous ovarian tumors. Using comparative genomic hybridization, we compared cytogenetic changes in borderline ovarian tumors and stage I invasive tumors. The average number of genetic alterations differed significantly between the borderline and the invasive tumors (1.9 and 9.2, respectively). The most common genetic alterations among the borderline tumors were loss of chromosome 17, 20q, and 18p, and gain of 12p13 approximately q23. These changes were also found among the invasive tumors in a similar percentage. In conclusion, we found four distinct cytogenetic alterations that might be early events in serous ovarian tumors, and that might also characterize a subgroup of borderline ovarian tumors that may have the potential to progress and develop malignancy.

Type-dependent E6/E7 mRNA expression of single and multiple high-risk human papillomavirus infections in cervical neoplasia.

BACKGROUND: Coinfection with multiple HPV types is common in cervical lesions, but the biological significance of the individual infections is difficult to establish. Expression of oncogenic E6/E7 HPV mRNA is correlated to risk of malignant progression, commercial assays for genotyping E6/E7 mRNA of all HR-HPV are lacking. OBJECTIVES: To characterize the tendency of 12 HR-HPV to express mRNA, correlated to the severity of the cervical lesion. Furthermore, we wanted to analyse mRNA expression in multiple infections, in order to establish which genotype may be responsible for cellular transformation. STUDY DESIGN: 245 samples from women with normal histology, various grades of dysplasia (cervical intraepithelial neoplasia grade 1-3), and cancer, were analysed for presence and genotyping of HPV DNA and mRNA using an in house real-time PCR test. RESULTS: Presence of mRNA was detected for 64% of the in total 422 HPV infections present in the samples, and more commonly in high-grade lesions. In 88% of DNA-positive samples from CIN2+ lesions, mRNA could be detected, compared to 33% of DNA-positive samples from women in screening with normal cytology. The genotype most prone to express mRNA in high-grade lesions was HPV45, followed by HPV16 and HPV31, less prone was HPV59. Expression of mRNA was significantly enhanced in CIN2+ lesions, an association also found for HPV16. In 52% of multiple infections (in which mRNA expression was generally more common), more than one genotype expressed mRNA, a phenomenon increasing with severity of lesion. Presence of mRNA could more often be detected in samples with multiple infections than in samples with single infections. CONCLUSIONS: The frequent expression of E6/E7 by HPV45 may promote oncogenicity and could be of clinical importance. Since presence of E6/E7 mRNA was common in multiple infections regardless of histology, multiple infection could be a clinically important finding. In multiple HPV infections, mRNA testing may identify the genotype that causes transformation. However, since mRNA expression of several genotypes in one sample is common, further and larger studies using complementary techniques are required.

Improved survival with clinical guidelines? Evaluation of a quality register linked to clinical guidelines for ovarian cancer in the western health care region in Sweden between 1 September 1993 and 1 June 1998.

BACKGROUND: New clinical guidelines (CGs) for ovarian cancer in the western health care region in Sweden were established, beginning in September 1993 and still in effect. METHODS: A retrospective evaluation of 5 years of quality registration linked to CGs for ovarian cancer in this region was undertaken. The study material comprised 718 patients. Relative survival rates for the studied patients were compared with National Cancer Register data for the western health care region during the same period. The National Cancer Register data were also used to compare survival rate during the studied period and the preceding 5-year period. RESULTS: Relative 5-year survival rate in our material was 46.1%. Relative survival in western Sweden during the studied period was found to be improved compared with that during the preceding period (P<0.02). CONCLUSIONS: The CGs have led to an improved, tighter organization, with fewer clinicians in special 'tumor teams' performing more aggressive tumor reduction surgery. Chemotherapy prescription is centralized, while the actual administration is decentralized. This has probably been important for the good 5-year survival results.