HPV prevalence and genotype distribution in 2,306 patients with cervical squamous cell carcinoma in central and eastern China.

Background: To explore the positivity rate and genotype distribution of human papillomavirus (HPV) in cervical squamous cell carcinoma (CSCC) tissues in central and eastern China and to provide theoretical basis for cervical cancer screening and prophylactic HPV vaccine development in China. Methods: DNA was extracted from paraffin-embedded tissues of CSCC samples and exfoliated cervical cells of cervical cancer screening populations. 23 HPV genotypes were detected by combining polymerase chain reaction (PCR) and reverse dot hybridized gene chip detection technology in 2,306 CSCC tissues and 10,245 cervical cancer screening populations. The genotype distribution of HPV infection was analyzed. Results: The overall infection rate of HPVs in 2,306 CSCC patients was 92.71%. The frequency of single-type HPV infection and multiple-type HPV infection were 86.48% and 13.51%, respectively. The most common HPV genotypes detected in Chinese CSCC tissues were HPV-16, HPV-18, HPV-31, HPV-33, HPV-45, HPV-52, HPV-58, and HPV-59. The overall positivity rate of these eight high-risk HPV (HR-HPV) genotypes in HPV-positive CSCC was as high as 96.91%. Of which the positivity rate of seven HR-HPV genotypes related to nine-valent HPV vaccines in HPV-positive CSCC was 95.09%. Meanwhile, the overall infection rates of HR-HPV and low-risk HPV (LR-HPV) in female aged 35-64 years who underwent cervical cancer screening were 13.16% and 1.32%, respectively. The high-frequency HR-HPV genotypes in cervical cancer screening women were HPV-52, HPV-58, HPV-16, HPV-53, HPV-68, HPV-39, HPV-51, and HPV-56, with positivity rates of 2.25%, 1.60%, 1.31%, 1.22%, 0.93%, 0.92%, 0.78%, and 0.74%, respectively. Conclusion: Among women screened for cervical cancer in China, detecting the 8 high-frequency HR-HPV genotypes can reduce technical difficulty and reagent costs, while also improving the efficiency and effectiveness of cervical cancer screening. HPV genotyping assists gynecologists in assessing the risk of HR-HPV-positive cervical intraepithelial neoplasia and guiding them in implementing appropriate interventions. Furthermore, HPV genotyping is helpful for doctors to follow up HR-HPV-positive women and to evaluate the protective effect of HPV vaccine.

Comparative Study on Detection and Typing of Human Papillomavirus (HPV) Infection with Microarray Using Paraffin-Embedded Specimens from Squamous Cell Carcinoma and Cervical Precursor Lesions.

Prevalence and type distribution of human papillomavirus (HPV) in squamous cell carcinoma (SCC) and cervical intraepithelial neoplasia (CIN) were investigated in Jiangsu Province, China. A total of 344 tissue sample cases with cervical squamous-cell carcinoma (SCC) and 135 with CIN I, 117 with CIN II, and 220 with CIN III were obtained from women undergoing biopsy or surgery. 33 samples of normal cervical tissue were collected with biopsy in inspected women. HPV detection and typing were examined by polymerase chain reaction (PCR) and gene-chip. Results showed that about 15.99% of cervical squamous-cell carcinoma, 34.09% of CIN III, 27.35% of CIN II and 10.37% of CIN I specimens exhibited multiple infections. Moreover, all total positive rate, single infection and multiple infection rates for the CIN I, CIN II, CIN III and SCC groups had significant difference (P < 0.05) when compared with the normal tissue group. The total positive rate, single infection and multiple infection rates for the CIN I, CIN II and CIN III groups showed significant difference (P < 0.05) when compared with the SCC group. All the CIN II, CIN III and SCC had intimate relationship with HPV infection. The high prevalence of HPV type 16, 18, 31, 33, 52, 58 and 59 in the cervical SCC deserves more attention in Jiangsu province.

Clinical Study of Styping Detection of Human Papillomavirus (HPV) Infection with Microarray from Paraffinembedded Specimens of Cervical Cancer and Precursor Lesions.

The prevalence and type distribution of human papillomavirus (HPV) in cervical cancer and cervical intraepithelial neoplasia (CIN) in Jiangsu, China was investigated. A total of 93 cases with cervical cancer and 176 CINII-III tissue samples were obtained from women undergoing biopsy or surgery. The 1047 exfoliated cervical cell samples were collected with cervical brush in physical examination women. HPV DNA and typing were examined by polymerase chain reaction (PCR) and gene-chip. The results showed that HPV DNA was detected in 82 cases with cervical cancer (88.17%), HPV 16 being detected in 65 (69.89%) cases, HPV 18 in 12 (12.90%) cases, HPV 33 in 10 (10.75%) cases, HPV 31 in 4 (4.30%) cases, and HPV 45 in 3 (3.23%) cases. HPV DNA was detected in 154 cases with CINII-III (87.50%), HPV 16 being detected in 92 (52.27%) cases, HPV 18 in 50 (28.41%) cases, HPV 33 in 25 (14.21%) cases, HPV 58 in 25 (14.21%) cases, and HPV 31 in 20 (11.36%) cases. About 20.43% cervical cancer and 38.64% CINII-III specimens exhibited multiple infections (p < 0.01). The total positive rate, single infection and mixed infection rate of the CINII-III and SCC group all had a significant difference (p < 0.05) when compared with the normal cells group. The total positive rate, single infection rate and mixed infection rate of CINII-III group did not show significant difference (p > 0.05) when compared with SCC group. CINII-III and SCC had all intimate relation with HPV infection. The high prevalence of HPV 16, 18, 33, 31 and 58 in Jiangsu (China) deserves more attention, as it has important implications for the successful use of HPV vaccine and choice of diagnostic methods.

[Analysis on human papillomavirus 16 and 18 types infection among 805 patients with common anorectal lesions].

OBJECTIVE: To analyze the infection condition of human papillomavirus (HPV) type 16 and 18 in the squamous cells and columnar cells of patients with common anorecatal lesions. METHODS: Infections of HPV type 16 and 18 were determined with real-time fluorescent quantitative PCR in the wax-embedded surgical specimen of 805 patients with common anorectal diseases. RESULTS: The overall infection rate among 805 patients with anorecatal lesions was 66.1% (532/805). The infection rate was 82.6% (95/115) in patients with mixed hemorrhoids, 76.5% (88/115) in anal papillary fibromas, 74.8% (86/115) in internal hemorrhoids, 72.2% (83/115) in fistulas, 69.6% (80/115) in external hemorrhoids, 47.8% (55/115) in anal perianal abscesses, and 39.1% (45/115) in anal fissures. CONCLUSION: Infection rate of HPV type 16, 18 in common anorectal lesions is high.

[Anticancer effects of tea polyphenols on colorectal cancer with microsatellite instability in nude mice].

OBJECTIVE: To study the anticancer effects of tea polyphenols on colorectal cancer with microsatellite instability (MSI) in nude mice and to explore its mechanism. METHODS: A colostomy was performed on the caecum of nude mice. Tumor fragments collected from the subcutaneous tumor of hMSH2-absence colon carcinoma Lovo cell line were surgically implanted onto the submucosa of the caecum during colostomy to establish the model. Then, the nude mice were divided into untreated group and 50, 75 and 100 mg/kg tea polyphenols groups. The mice in tea polyphenols-treated groups were given intra-abdominal injection of 50, 75 and 100 mg/kg tea polyphenols respectively. The inhibition rates of tumors were calculated, and microsatellite instability (MSI) and the alteration of transforming growth factor-beta1 (TGF-beta1), TGF-beta2 and insulin-like growth factor (IGF) were detected by Genescan method at different times after the injection. RESULTS: The tumor volumes of the three groups began to decrease at the 1st week and decreased most greatly from 2 to 3 weeks after treatment, and then the tumors tended to increase. The study found that tea polyphenols could inhibit the tumor growth. The tumor inhibition rates in the three treated groups were significantly higher than those in untreated group 1, 2, 3 and 4 weeks after treatment (P<0.01). Detection of MSI showed that the colorectal tumor in the untreated group presented with four MSI signs, including BAT-25, D2S123, D5S346 and D17S250, and TGF-beta1, TGF-beta2, IGF expressions. After using the tea polyphenols, the microsatellite tended to become stable. CONCLUSION: Tea polyphenols can inhibit the mismatch-repair-gene deficient colorectal cancer in nude mice by down-regulating the microsatellite instability.

Detection of mismatch repair gene germline mutation carrier among Chinese population with colorectal cancer.

BACKGROUND: Hereditary nonpolyposis colorectal cancer (HNPCC) is an autosomal dominant syndrome. The National Cancer Institute (NCI) has recommended the Revised Bethesda guidelines for screening HNPCC. There has been a great deal of research on the value of these tests in other countries. However, literature about the Chinese population is scarce. Our objective is to detect and study microsatellite instability (MSI) and mismatch repair (MMR) gene germline mutation carriers among a Chinese population with colorectal cancer. METHODS: In 146 prospectively recruited consecutive patients with clinically proven colorectal cancer, MSI carriers were identified by analysis of tumor tissue using multiplex fluorescence polymerase chain reaction (PCR) using the NCI recommended panel and classified into microsatellite instability-low (MSI-L), microsatellite instability-high (MSI-H) and microsatellite stable (MSS) groups. Immunohistochemical staining for MSH2, MSH6 and MLH1 on tissue microarrays (TMAs) was performed, and methylation of the MLH1 promoter was analyzed by quantitative methylation specific PCR (MSP). Germline mutation analysis of blood samples was performed for MSH2, MSH6 and MLH1 genes. RESULTS: Thirty-four out of the 146 colorectal cancers (CRCs, 23.2%) were MSI, including 19 MSI-H CRCs and 15 MSI-L CRCS. Negative staining for MSH2 was found in 8 CRCs, negative staining for MSH6 was found in 6 CRCs. One MSI-H CRC was negative for both MSH6 and MSH2. Seventeen CRCs stained negatively for MLH1. MLH1 promoter methylation was determined in 34 MSI CRCs. Hypermethylation of the MLH1 promoter occurred in 14 (73.7%) out of 19 MSI-H CRCs and 5 (33.3%) out of 15 MSI-L CRCs. Among the 34 MSI carriers and one MSS CRC with MLH1 negative staining, 8 had a MMR gene germline mutation, which accounted for 23.5% of all MSI colorectal cancers and 5.5% of all the colorectal cancers. Five patients harbored MSH2 germline mutations, and three patients harbored MSH6 germline mutations. None of the patients had an MLH1 mutation. Mutations were commonly located in exon 7 and 12 of MSH2 and exon 5 of MSH6. Right colonic lesions and mucinous carcinoma were not common in MSI carriers. CONCLUSION: Our data may imply that the characteristics of HNPCC in the Chinese population are probably different from those of Western countries. Application of NCI recommended criteria may not be effective enough to identify Chinese HNPCC families. Further studies are necessary to echo or refute our results so as to make the NCI recommendation more universally applicable.

[Study the value of screening hereditary nonpolyposis colorectal cancer kindreds by detecting the expression of hMLH1/hMSH2 with tissue microarray].

OBJECTIVE: To study the value of screening hereditary nonpolyposis colorectal cancer (HNPCC) kindreds by detecting the expressions of hMLH1/hMSH2 with tissue microarray. METHODS: A tissue microarray with 22 colorectal cancers from HNPCC families and 15 sporadic colorectal cancers was established, and the expressions of hMLH1/hMSH2 were detected by immunohistochemistry (IHC). RESULTS: The expressions of hMLH1 or hMSH2 were negative in 15 of 22 HNPCC and 1 of 15 sporadic colorectal cancers in routine IHC. The expressions of hMLH1 or hMSH2 were negative in 17 of 22 HNPCC and 2 of 15 sporadic colorectal cancers in tissue microarray. The examination of hMSH2 expression yielded same results between routine IHC and tissue microarray. There were no difference on the hMLH1 expressions between routine IHC and tissue microarray. CONCLUSION: Tissue microarray is a high-throughput way to detect the expressions of hMLH1/hMSH2 and is applicable to screen HNPCC kindreds.