Huaizeng Chen

Stem-cell-abundant proteins Nanog, Nucleostemin and Musashi1 are highly expressed in malignant cervical epithelial cells

BackgroundNanog, nucleostemin (NS) and musashi1 (Msi1) are proteins that are highly expressed in undifferentiated embryonic stem (ES) cells and have been shown to be essential in maintaining the pluripotency and regulating the proliferation and asymmetric division of ES cells and several nervous system tumor cells. The roles of Nanog, NS and Msi1 in development and progression of cervical carcinoma have, until now, not been well documented.MethodsIn this study, expression of Nanog, NS and Msi1 was detected by immunohistochemistry analysis in 235 patients with various degrees of cervical epithelial lesions, including 49 with normal cervical epithelia, 31 with mild dysplasia (CIN I), 77 with moderate-severe dysplasia (CIN II-III) and 78 with squamous cervical carcinomas (SCCs). Associations with various clinical pathological prognostic variables were analyzed in 50 early-stage SCC patients.ResultsNanog, NS and Msi1 expression levels were significantly higher in SCC patients compared with CIN patients, and were higher in CIN patients compared with those with normal cervical epithelia. Nanog expression levels showed significantly differences according to different tumor sizes (P < 0.05), whereas there were no differences in NS and Msi1 expression levels according to different clinical pathological parameters.ConclusionOur findings indicate that Nanog, NS and Msi1 may be involved in carcinogenesis of the cervix and progression of cervical carcinoma.

Proportion of CD4+CD25+ Regulatory T Cell is Increased in the Patients with Ovarian Carcinoma

Objective: To study the frequency of the CD4+CD25+ regulatory T cells (Tregs) in the patients with ovarian carcinoma and its possible mechanism. Methods: The percentages of CD4+CD25+ Tregs in the peripheral blood lymphocytes (PBLs), tumor infiltrating lymphocytes (TILs) and tumor associated lymphocytes (TALs) from 13 patients with ovarian carcinoma and in the PBLs from 14 healthy women were determined by flow cytometry. The expression of CD69 on CD4+PBLs from the patients was detected. PBLs from healthy women were cultured in complete RPMI 1640 containing the supernatant from SKOV3 cell line with or without PHA (phytohemagglutinin) stimulation for 72 hours, then the percentage of CD4+CD25+ T cells was detected. Results: CD4+CD25+ Tregs in the PBLs from patients with ovarian carcinoma were significantly increased compared with those from the control. The percentage of CD4+CD25+ Tregs in TILs was higher than that in PBLs and TALs from the patients, but not significantly. The expression of CD69 on CD4+PBLs from the patients was negative. The percentages of CD4+CD25+ T cells in PBLs cultured with SKOV3 supernatant elevated significantly compared with those without supernatant whether PHA was added or not (P = 0.001 and 0.001, respectively). Conclusion: There is an increasing of the proportion of CD4+CD25+ Tregs in PBLs, TILs and TALs of the patients with ovarian carcinoma, which probably results from up-regulation of soluble factor secreted by ovarian carcinoma cells.

Expression of Sox2 in human ovarian epithelial carcinoma

ObjectivesThe aim of this study was to investigate the expression of Sox2, a transcription factor, in a series of benign, borderline, and malignant ovarian tumors and to evaluate whether Sox2 expression levels correlate with clinicopathological characteristics in ovarian epithelial carcinoma.MethodsThis study investigated immunohistochemical expressions of Sox2 in 43 species of normal ovarian epithelia, 284 species of serous epithelial lesions, and 164 species of mucinous epithelial lesions to assess their clinicopathological relevance.ResultsImmunohistochemical results showed that the positive ratio of Sox2 expression gradually increased from benign and borderline to malignant ovarian tumors; 55.81% of normal ovarian epithelia, ~65% of serous and mucinous cystadenoma, ~70% of borderline serous and mucinous cystadenoma, and ~91% of serous and mucinous cystadenocarcinoma expressed Sox2, respectively. However, there was no significant correlation between Sox2 expression and the age and level of CA125 in patients with either serous or mucinous tumors. Positive correlations between Sox2 expression levels and FIGO stage or pathological stage were identified in both serous and mucinous cystadenocarcinoma samples.ConclusionThe expression level of Sox2 in human ovarian tumors was directly proportional to their degree of malignancy, implying that Sox2 overexpression may be closely related to the malignant transformation of ovarian tumors.

Nanog Is Highly Expressed in Ovarian Serous Cystadenocarcinoma and Correlated with Clinical Stage and Pathological Grade

Objective: Nanog is overexpressed in embryonic stem cells for cell self-renewal and differentiation. We investigated whether the Nanog expression is associated with the occurrence and development of ovarian cancer. Methods: Immunohistochemistry was used to examine the expression of Nanog in 43 normal ovarian epithelia, 110 serous cystadenomas, 80 borderline serous cystadenomas, and 107 serous cystadenocarcinomas. In the meantime, their association with various clinicopathologic features was assessed. Results: The expression intensity of Nanog in normal ovarian tissue, benign, borderline, and malignant tumors showed a gradual rising trend. Among the serous cystadenocarcinomas, 42.86% were detected to be positive for stage I, 70.97% for stage II, 95.31% for stage III, and 100% for stage IV. There was a strong correlation between Nanog and clinical stage (r = 0.418, p = 0.000). Besides, there was a 55.56% positive expression of grade I, 73.68% of grade II, and 96.67% of grade III. The correlation between Nanog and differentiation grade was dramatic (r = 0.692, p = 0.000). Conclusions: Nanog was highly expressed in ovarian serous cystadenocarcinoma, and showed a positive correlation with clinical stage and grade. Nanog may play an important role in the development of dedifferentiation and progression of serous ovarian carcinoma.

The Expression of Interleukin-10 in Patients with Primary Ovarian Epithelial Carcinoma and in Ovarian Carcinoma Cell Lines:

This study determined interleukin-10 (IL-10) expression in patients with ovarian carcinoma and in ovarian carcinoma cell lines, and investigated its clinical significance in the development and progression of ovarian carcinoma. Expression of IL-10 in ovarian carcinoma, benign ovarian tumour, normal control tissues and ovarian carcinoma cell lines was detected by immunohistochemistry and Western blot analysis. IL-10 concentrations in sera and ascites from patients with ovarian carcinoma, in sera from patients with benign ovarian tumour and normal controls, and in supernatants of ovarian carcinoma cell line cultures were measured by enzyme-linked immunosorbent assay. The tissue level of IL-10 in ovarian carcinoma was significantly higher than in benign ovarian tumour and normal controls. IL-10 expression was detectable in cell lysate and supernatant from ovarian carcinoma cell lines. In patients with ovarian carcinoma the IL-10 level in ascitic fluid was significantly higher than in sera, and the serum IL-10 level in ovarian carcinoma was significantly higher than in benign ovarian tumour and normal controls. Ascitic IL-10 levels in ovarian carcinoma were significantly correlated with disease stage but not cytological grade. These results suggest that ovarian carcinoma cells are able to synthesize and secrete IL-10, which probably assists in promoting the development and progression of ovarian carcinoma.

Susceptibility of XRCC3, XPD, and XPG Genetic Variants to Cervical Carcinoma

Objective: DNA repair genes play a key role in maintaining genomic stability and integrity. DNA repair gene polymorphisms, such as those of XRCC3 and xeroderma pigmentosum, complementation group D and G (XPD, XPG), contribute to carcinogenesis. In this study, we investigated the correlation between cervical carcinoma risk and XRCC3, XPD, XPG genetic variants. Methods: A case-control study of 400 cases including 200 carcinoma, 200 cervical intraepithelial neoplasia (CIN) and 200 normal women was performed. Four single nucleotide polymorphisms (SNPs) (XRCC3 Thr241Met, XPG His1104Asp, XPD Asp312Asn, and XPD Lys751Gln) were genotyped by mismatch amplification polymerase chain reaction. Results: Women carrying homozygous Asp1104Asp genotypes had a significantly decreased risk of cervical or cervical squamous cell carcinoma compared to His1104Asp or His1104His genotypes. Similarly, XPD Asn312Asn (AA) reduced the risk of cervical or cervical squamous cell carcinoma. No association of XRCC3 Thr241Met or XPD Lys751Gln and cervical carcinoma was found. None of the 4 SNPs influenced the risk of CIN in our study. Conclusion: Our results support the hypothesis that genetic variations in DNA repair genes may contribute to an inherited genetic susceptibility to cervical carcinoma.

PARP-1 Val762Ala Polymorphism Is Associated with Risk of Cervical Carcinoma

PARP-1 is a nuclear enzyme that plays an important role in DNA repair, recombination, proliferation and the genome stability. The PARP-1 Val762Ala polymorphism has been associated with increased risk of developing cancers of the prostate, esophagus and lung. The aim of this study was to determine whether the PARP-1 Val762Ala polymorphism is associated with the risk of cervical carcinoma. MA-PCR was used to genotype the PARP-1 Val762Ala polymorphism in 539 women with cervical carcinoma, 480 women with CIN and 800 controls. The genotyping method was confirmed by the DNA sequencing analysis. The PARP-1 Val762Ala polymorphism was not associated with the risk of CIN. However, women carrying the PARP-1 Ala762Ala genotype were significantly susceptible to cervical carcinoma (OR: 2.70, 95% CI: 1.47–3.70), and the similar results were also found in squamous cell carcinoma (OR: 2.56, 95% CI: 1.47–3.70). In HPV positive population, the PARP-1 Ala762Ala genotype was also associated with increased risk of cervical carcinoma (OR: 5.56, 95% CI: 2.08–14.3). Our results indicate that the PARP-1 Ala762Ala genotype increases the risk of cervical carcinoma.

The effect of five probiotic lactobacilli strains on the growth and biofilm formation of Streptococcus mutans

OBJECTIVE To compare the effects of five probiotic lactobacilli strains on the growth and biofilm formation of Streptococcus mutans (MS). MATERIALS AND METHODS Five probiotic lactobacilli bacteria (LB), Lactobacillus casei Shirota, Lactobacillus casei LC01, Lactobacillus plantarum ST-III, Lactobacillus paracasei Lpc-37, and Lactobacillus rhamnosus HN001, were used as test strains effecting on the Streptococci strain S. mutans UA159 in this study. The effect of LB strains and their supernatants on the viability of the MS was evaluated. Then, the effect of LB strains on the growth of MS biofilm formation was observed by fluorescence microscope. RESULTS All of the LB strains inhibited the growth of MS at concentrations of 1 × 10(8) and 3 × 10(8) CFU ml(-1) (P < 0.05). Untreated (without pH adjustment and ultrafiltration) LB supernatants from all of the LB strains inhibited the growth of MS (P < 0.05) as well. After pH adjustment and ultrafiltration (treated), only supernatants from L. casei Shirota and L. rhamnosus HN001 inhibited the growth of MS (P < 0.05). MS biofilm formation was also inhibited by all untreated supernatants and by the treated supernatants of L. casei Shirota and L. rhamnosus HN001 (P < 0.05). CONCLUSION All five probiotic lactobacilli strains inhibited the growth and biofilm formation of MS, likely through the production of an acid environment, bacteriocin-like poly peptides, or both, and the effects on MS were dependent on the LB strains used.

Expression of octamer-4 in serous and mucinous ovarian carcinoma

Background Octamer-4 (Oct4) is a well known regulator of self-renewal in embryonic stem cells; it has been detected in several human cancers and may play a critical role in carcinogenesis. Aims To assess the expression of Oct4 in epithelial ovarian tumours. Methods Expression of Oct4 was evaluated by immunohistochemistry in 460 cases of various epithelial ovarian lesions as well as 35 cases of normal fallopian tube epithelium. The association between Oct4 expression and various clinical pathological parameters was analysed. Results Oct4 expression was significantly increased from normal epithelium (both ovarian epithelium and fallopian tube epithelium) to benign and borderline cystadenoma to carcinoma in the serous lesion subgroup. Oct4 overexpression was associated with more advanced FIGO stage and higher histological grade in serous adenocarcinoma. Conversely, Oct4 expression did not differ among mucinous lesions or correlate with clinicopathological parameters in patients with mucinous adenocarcinoma. Conclusion Results suggest that Oct4 expression may contribute to the initiation, promotion and progression of serous ovarian carcinoma; it might be a useful biomarker for the diagnosis and outcome prediction of serous ovarian carcinoma.

Hes1/Hes5 gene inhibits differentiation via down-regulating Hash1 and promotes proliferation in cervical carcinoma cells.

Introduction: Hairy and enhancer of split 1 (Hes1) and Hes5 are target genes for the mammalian Notch pathway, which are highly expressed in epithelia in the process of embryogenesis or in neural stem cells, inhibit cell differentiation via the Notch-Hes-Hash signaling, and promote the survival of stem cells. Either Hes1 or Hes5 overactivation is likely to affect cell differentiation, thereby resulting in carcinogenesis. Methods: We transfected the diced small interference RNA into SiHa cells and detected cell differentiation and proliferation by immunocytochemistry, Western blot, and methyl thiazolyl tetrazolium assay. Results: Knockdown of Hes1 and Hes5 would up-regulate the downstream gene Hash1, but not the upstream gene Notch1 in the Notch-Hes-Hash pathway. After Hes1/Hes5 RNA interference, expression of differentiation-associated proteins (including Nanog, stage-specific embryonic antigen 4, and tumor rejection antigen-1-60) was reduced, and the cell differentiation was promoted; meanwhile, the cell proliferation was inhibited, which was verified by detecting proliferation-associated proteins (eg, Ki-67, proliferating cell nuclear antigen) and methyl thiazolyl tetrazolium assay. Conclusions: Our findings suggest that Hes1/Hes5 gene would inhibit the cell differentiation via down-regulating Hash1 and promote the cell proliferation in cervical carcinoma cells; the cell differentiation and proliferation can be reversed simultaneously by Hes1/Hes5 knockdown using RNA interference.