Screening for MiRNAs related to laryngeal squamous carcinoma stem cell radiation.

OBJECTIVE: To use microarray chip technology for screening of stem cell radiation related miRNAs in laryngeal squamous cell carcinoma; study and explore the relationship of miRNAs with radiosensitivity of laryngeal squamous cells. METHOD: After conventional culture and amplification of the laryngeal squamous carcinoma cell line Hep-2, CD 133+ cells were screened out with combination of isolated culture of stem cell microspheres and FACS for preparation of laryngeal cancer stem cells. After radiation treatment, miRNAs of laryngeal squamous carcinoma stem cells before and after radiation were enriched and purified. After microarray hybridization with mammalian miRNA and scanning of fluorescence signal, the miRNAs of laryngeal squamous carcinoma stem cells before and after radiation was subject to differential screening and clustering analysis. Real-time quantitative RT-PCR was used to verify part of the differentially expressed miRNAs. RESULTS: 70 miRNAs related to laryngeal cancer stem cell radiation with 2-fold difference in expression were screened out, in which 62 were down-regulated and 8 were up-regulated. Fluorescent quantitative RT-PCR results were consistent with miRNAs chip results. CONCLUSION: Some miRNAs may be involved in self-regulation with laryngeal squamous carcinoma stem cell radiation.

Reduced expression of aquaporin 9 in tubal ectopic pregnancy.

Previous studies demonstrate significant roles for passive water channels (aquaporins, AQPs) in maintaining water homeostasis in cell membranes of endometrial cells during decidualisation and embryo implantation. However, there is little information regarding the role of AQPs in the human fallopian tube, specifically their role in human tubal ectopic pregnancy. In this study we took tissue samples from the site of implantation of tubal ectopic pregnancy (group 1, N = 30, mean age 32 years, range 23-42) and the corresponding non-implantation site in women undergoing salpingectomy for tubal pregnancy (group 2). Ampullary fallopian tubes during mid-secretory phase were collected as control group (group 3, N = 17, mean age 37 years, range 30-50). Thin sections were prepared and stained with anti-AQP9, and, for estrogen and progesterone receptors in each group. Immunohistochemical studies showed that AQP9 proteins localize in the cytoplasm of epithelial cells of Fallopian tube. Expression of AQP9 was significantly reduced during tubal pregnancy compared to controls (group 1 vs. group 3, P = 0.036; group 2 vs. group 3, P = 0.029), and, this reduced expression was not related to estrogen receptor or progesterone receptor status (group 2, ER vs. AQP9, Pearson r = 0.173, P = 0.361; PR vs. AQP9, Pearson r = 0.124, P = 0.514, respectively). Similarly, there is no correlation between AQP9 and estrogen receptor or progesterone receptor status in the normal group (group 3, ER vs. AQP9, Pearson r = -0.026, P = 0.923; PR vs. AQP9, Pearson r = -0.292, P = 0.255, respectively). Reduced expression of AQP9 in human fallopian tube may contribute to aspects of pathophysiology of tubal ectopic pregnancy.

Locally elevated leukemia inhibitory factor in the inflamed fallopian tube resembles that found in tubal pregnancy.

OBJECTIVE: To investigate the association between the expressions of leukemia inhibitory factor and the occurrence of tubal pregnancy. DESIGN: Prospective observational study. SETTING: University-based obstetrics and gynecology hospital. PATIENT(S): Thirty women undergoing salpingectomy for tubal pregnancy and 30 nonpregnant patients with benign uterine or appendix disease. INTERVENTION(S): Oviduct tissues with ectopic gestation were separated into implantation sites and nonimplantation sites. Samples of ampullary fallopian tubes during midsecretory phase were collected as control groups. Immunohistochemical and Western blot analysis were performed. MAIN OUTCOME MEASURE(S): The differences of leukemia inhibitory factor expression between the implantation and nonimplantation sites of oviduct tissues and the normal and chronically inflamed fallopian tubes. RESULT(S): The expression of leukemia inhibitory factor in the implantation group is significantly higher than that in the nonimplantation group or in the normal group. A statistically significant difference was also found for leukemia inhibitory factor between the chronic inflammation group and the normal group by Western blot analysis but no difference between the chronic inflammation group and the implantation group or the nonimplantation group. CONCLUSION(S): Leukemia inhibitory factor might be one of the reasons that cause patients with salpingitis to be more susceptible to tubal pregnancy and might be involved in the implantation process of tubal pregnancy.

[Expression of aquaporin-8 and bcl-2 protein in human cervical carcinoma and their correlations].

OBJECTIVE: To investigate the expression of aquaporin-8 (AQP8) and apoptosis associated bcl-2 protein in human cervical carcinoma and their relationship. METHODS: The expression of AQP8 and bcl-2 protein in 74 cases of cervical carcinoma (46 cases of squamous-cell carcinoma of the uterine cervix, 28 cases of adenocarcinoma of the uterine cervix), 34 cases of cervical intraepithelial neoplasia (CIN) and 15 cases of normal cervices were detected by immunohistochemical technique, and their clinical significance were analyzed. RESULTS: The expression of AQP8 and bcl-2 protein were detected in intracytoplasm of atypia cells in CIN, squamous-cell carcinoma and adenocarcinoma of the uterine cervix. The positive rates of AQP8 and bcl-2 in squamous-cell carcinoma, adenocarcinoma, CIN and normal cervical epithelium were 98%, 74%; 61%, 71%; 71%, 53% ; 53%, 20% respectively. There were significant differences between squamous-cell carcinoma of the uterine cervix and other groups in AQP8 (P < 0.01), but no significant differences were found in any other groups. There were significant differences between squamous-cell carcinoma of the uterine cervix and CIN or normal cervical epithelium in bcl-2, so were between adenocarcinoma of the uterine cervix. The expression of AQP8 was positively correlated with bcl-2 in human cervical carcinoma( r(s) = 0.463, P = 0.000). CONCLUSIONS: There is a close relationship between high expression of AQP8 and development of human cervical carcinoma. The expression of AQP8 protein is positively correlated with bcl-2 protein in human cervical carcinoma. AQP8 protein may have anti-apoptosis function, although the detailed mechanism in human cervical carcinoma remains to be clarified.

[Microvessel density, epithelial-stromal vascular cuffing and expression of vascular endothelial growth factor in human cervical carcinoma].

OBJECTIVE: To observe microvessel density(MVD), epithelial stromal vascular cuffing(VC) and expression of vascular endothelial growth factor(VEGF) in human cervical carcinomas and to clarify their significance in the invasion and metastasis of cervical carcinoma. METHODS: VEGF and CD34 were stained immunohistochemically (SP) in 57 cases of cervical carcinoma (30 cases of squamous cell carcinoma, 20 of adenocarcinoma 7 of glandular and squamous cell carcinoma), 29 cases of cervical intraepithelial neoplasia (CIN) and 16 cases of normal cervices, meanwhile, MVD and VC were also assayed. RESULTS: There were significant differences among the above 5 groups for MVD P<0.01 . The VC pattern showed a significant difference between cervical carcinoma and CIN or control group P<0.01). The positive rates of VEGF in normal cervical epithelium, CIN, squamous cell carcinoma, adenocarcinoma, glandular and squamous cell carcinoma were 18.8% 3/16, 82.8% 24/29), 93.3% 28/30), 100% 20/20 and 7/7(100%), respectively. There were significant differences between these cervical lesion groups and the control group(P<0.001). The MVD showed significant differences between the positive pelvic node metastasis and negative pelvic node metastasis P<0.05). There was no significant correlation between the expression of VEGF and the tumor diameter, clinical stage, pathologic grade and pelvic node metastasis. CONCLUSION: The expression of VEGF may play an important role in the angiogenesis of cervical carcinoma. Degree of malignancy of cervical carcinoma has a close association with microvessel density.

[Expression of matrix metalloproteinase-2, 9 and their inhibitor-TIMP 1,2 in human squamous cell carcinoma of uterine cervix].

BACKGROUND & OBJECTIVE: Studies of tumor invasion and metastasis have focused on the degradation of extracellular matrix (ECM). Matrix metalloproteinases (MMPs), which could degrade ECM, were implicated in cancer invasion and metastasis. Activated MMPs were controlled by specific tissue inhibitors of metalloproteinases (TIMPs). This study was designed to investigate the expression of MMP-2, MMP-9 and TIMP-1, TIMP-2 in human squamous cell carcinomas of uterine cervix, and the association with invasion and metastasis of human squamous cell carcinoma of uterine cervix. METHODS: Tissue samples from 40 cases of squamous cell carcinoma of the uterine cervix, 29 cases of cervical intraepithelial neoplasia (CIN), and 16 cases of normal cervices were stained immunohistochemically. RESULTS: Immunohistochemical staining of tumor cells for MMP-2, MMP-9, TIMP-1, and TIMP-2 were noted 82.5%, 70.0%, 62.5%, and 97.5% in carcinomas, 44.8%, 62.1%, 86.21%, and 96.55% in CIN lesions respectively, for MMP-2, MMP-9, TIMP-1 in 25.0% normal cervices, but for TIMP-2 in 56.25% normal cervices. There was significant difference for MMP-2 among carcinomas, normal cervices, and CIN respectively (P < 0.01). There was significant difference for MMP-9 among normal cervices, carcinomas, and CIN, respectively (P < 0.05). The positive rates of TIMP-1 and TIMP-2 in the cervical carcinoma group and CIN group were obviously higher than that in normal cervical group(P < 0.01). The positive rates of MMP-2 and MMP-9 in lymph node metastasis were obviously higher than that in without lymph node metastasis(P < 0.01), but for TIMP-2 was contrary and there was no significant difference for TIMP-1. No significant correlation could be established among the expression of these markers and the tumor diameter, the clinical stage, and pathologic grade. CONCLUSIONS: The over-expression of MMP-2, MMP-9, TIMP-1, and TIMP-2 may play a key role in invasion and lymph-node metastasis of in squamous carcinoma of the cervix.