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The Roles of Hedgehog Pathway in Cervical Carcinoma

Author: LiGuiLing
Tutor: ChenZhengAi
School: Yanbian University
Course: Pharmacology
Keywords: Hh signaling pathway Cervical Cancer cyclopamine
CLC: R737.33
Type: Master's thesis
Year: 2008
Downloads: 79
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Abstract


Objective: To investigate the Hedgehog (Hh) signaling pathway in human cervical carcinoma cell lines and expression, and preliminary study Hh signaling pathway inhibitor cyclopamine on cervical cancer cell line HeLa and SiHa cytotoxicity and mechanisms for the early diagnosis of cervical cancer and The discovery of new therapeutic targets provide a theoretical basis. Methods: (1) by immunohistochemical staining Shh, Ihh, Ptch, Smo, Gli-1, Gli-2, Gli-3 antibody in 38 cases of squamous cell carcinoma, 23 cases of CIN Ⅲ, 10 例 CIN Ⅱ, 4 例CIN Ⅰ, 32 cases of normal cervical epithelial tissue expression and distribution; (2) extract 38 cases of cervical cancer DNA, using polymerase chain reaction (PCR) detection of human papillomavirus (Human Papillomavirus, HPV) 16 and 18; (3) MTT assay cyclopamine on cultured HeLa and SiHa cervical cancer cell line growth inhibition, while observing cellular morphological changes; (4) by means of immunocytochemical staining Hh signaling pathway in HeLa cells related genes and proteins Bax and Bel-2 expression. Results: (1) in cervical cancer and precancerous lesions (CIN Ⅱ / Ⅲ) in Hh signaling pathway related gene expression were significantly higher than in normal cervical mucosa. In addition to Gli-1/2 expression in the cytoplasm and nucleus, the rest of Hh signaling pathway related gene expression in the cytoplasm. Ihh, Ptch, Smo, Gli-1, Gli-3 in normal cervical epithelium almost no expression. In cervical cancer, the seven kinds of Hh signaling pathway gene expression compared with normal cervical tissue significantly increased (P <0.05). Shh expression in normal cervical epithelium to CIN CIN to cervical changes and transformation process was double (both) increased. These results suggest that Hh signaling pathway in cervical cancer and CIN is widely activated. (2) 38 cases of HPV16-positive rate was 71.1%, while HPV18 were negative. HPV16 in cervical cancer and 18 infected with Hh signaling pathway gene expression was no significant relationship. (3) MTT colorimetric assay showed that cyclopamine inhibits proliferation of HeLa and SiHa cells, the inhibition rate showed a dose-dependent manner; observed under an inverted microscope, two cell lines cyclopamine group appeared on both the shape and the number of changes. (4) immunocytochemistry showed by cyclopamine-treated HeLa cells Hh signaling pathway gene protein and Bcl-2 expression levels than the control group (tomatidine) was significantly lower; while Bax expression levels increased. Conclusions: Hh signaling pathway activation in cervical abnormalities, and participate in the development and progression of cervical cancer; this pathway inhibitor cyclopamine-induced apoptosis through inhibition of cancer cell proliferation. Hh signaling pathway may therefore be regarded as a new target for cancer therapy.

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CLC: > Medicine, health > Oncology > Genitourinary tumors > Female genital tumors > Uterine tumors
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