| Prostate cancer is one of the most prevalent malignant tumors threaten male health. It is the most common form of non-cutaneous cancer and second most lethal cancer in American men with an incidence of 232,090 new cases, and 30,350 deaths in 2005 alone. The incidence rate of prostate cancer in China is going up recently, and reported cases already up to the third in urinary malignant tumor. Although in many cases this cancer is treatable through hormone therapy and/or surgery, these cures are associated with a host of medical problem(e.g. lipid metabolic disturbance and heart diseases). And often provide only temporary relief, as prostate cancer reoccurs in part of cases as a far less treatable and much more malignant disease-androgen independent prostate cancer(AIPC).Recent studies showed high fat/high cholesterol'Western-type diets'have been linked to PCa incidence and progression. High levels of serum cholesterol contribute to prostate cancer progression and upon stimulation with androgen. Modification of cholesterol levels through diet, exercise and the use of pharmaceuticals may contribute to slowing prostate cancer Progression.In our studies we demonstrate that Hedgehog protein expression decreased after treatment in cholesterol depletion media (CDM) and the tumor cells apoptosis ratio increased. Hedgehog gene is a sub-segment polarity genes, it is named because that in Drosophila embryos showed hairball-like which resembles a frightened Hedgehog to follow gene mutations. There are three Hedgehog homologous genes in mammalian: Sonic Hedgehog (SHH), Indian Hedgehog (IHH) and Desert Hedgehog (DHH), encoding Shh, Ihh, and Dhh proteins respectively. Hedgehog have important physiological and pathological significance in cell differentiation, embryonic development, organ formation, injury repairs and tumorigenesis, but the specific molecular mechanism has not been fully clarified about cholesterol promoting the apoptosis of prostate cancer. Whether or not Hedgehog is the major cholesterol-sensitive factors which affect the prostate cancer cell proliferation, as well as low-cholesterol conditions, whether it is inhibited by Hedgehog downward effect on the proliferation of prostate cancer there were no relevant reports. Our study research the prostate cancer PC-3 cells, LNCaP cells and LNCaP-AI cell proliferation, apoptosis by MTT experiments, and the prostate cancer cells were cultured in normal medium (NM), cholesterol depletion of culture medium (CDM), high cholesterol medium (HCM) and the addition of simvastatin in each group, then we use semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR), Real-time PCR, ELISA method to detect the mRNA and protein levels of Hedgehog change; the same time Detection of Hedgehog signaling pathway downstream of protein Ptc, Smo and Gli expression of the mRNA; and last we use confocal laser scanning detection of sub-cellular localization of Hedgehog; to determine the low cholesterol and lipid-lowering (simvastatin) conditions, whether inhibited Hedgehog promotes cell apoptosis and inhibit prostate cancer cell from hormonal-dependent to-independent transformation, thus demonstrated that the Hedgehog is a cholesterol-sensitive cytokine hypothesis in prostate cancer cells. Revealed the fact that low-cholesterol can down-regulating the expression of Hedgehog, then inhibits the development of prostate cancer, provide a new way of thinking for the treatment of prostate cancer.Objective:To comparative study the proliferation, apoptosis as well as differences in the expression of Hedgehog signaling pathway of human prostate cancer PC-3 cells, LNCaP cells and LNCaP-AI cell which were cultured in NM, CDM, HCM groups, To analyze the impact of cholesterol on Hedgehog signaling pathway and prostate cancer progression.Method:Human prostate cancer PC-3 cells, LNCaP cells and LNCaP-AI cells were cultured in NM, CDM, and in HCM. MTT method to detect cell proliferation and apoptosis, using semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR), Real-time PCR, ELISA method to detect the change of Hedgehog mRNA and protein levels; Simultaneous detect Hedgehog signaling pathway protein Ptc, Smo and Glil; and observe the cell location of Hedgehog under the conditions of different concentrations of cholesterol by confocal laser.Result: 1, the growth curves of prostate cancer cells cultured in CDM were significantly lower than cultured in NM and HCM cells.2, the Hedgehog protein levels of prostate cancer cells cultured in CDM were significantly lower than that in NM and HCM, it is a significant difference.3, Hedgehog signaling pathway-related protein of prostate cancer cells cultured in CDM, the expression of Hedgehog, Smo and Gli increased and Ptc expression reduced.4, we also observed that the CDM group of prostate cancer cells express Hedgehog protein in cells was significantly higher than NM and HCM groups by laser confocal.Conclusions:1, low-cholesterol inhibits prostate cancer PC-3 cells, LNCaP cells and LNCaP-AI cells in vitro.2, low-cholesterol inhibits Hedgehog protein in the process of extracellular secretion of prostate cancer PC-3 cells, LNCaP cells and LNCaP-AI cells.3, low-cholesterol inhibit prostate cancer cell proliferation correlate with the fall of extracellular secretion of Hedgehog.Objective:To comparative study the proliferation, apoptosis as well as differences in the expression of Hedgehog signaling pathway of human prostate cancer PC-3 cells, LNCaP cells and LNCaP-AI cell which were cultured in NM, HCM with different concentration of Simvastatin, To analyze the impact of Simvastatin on Hedgehog signaling pathway and prostate cancer progression.Method:Human prostate cancer PC-3 cells, LNCaP cells and LNCaP-AI cells were cultured in NM, and in HCM, each group plus different concentrations of simvastatin. MTT method to detect cell proliferation and apoptosis, using semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR), Real-time PCR, ELISA method to detect the change of Hedgehog mRNA and protein levels; Simultaneous detect Hedgehog signaling pathway protein Ptc, Smo and Gli1; and observe the cell location of Hedgehog under the conditions of different concentrations of simvastatin by confocal laser.Result:1, the growth curves of prostate cancer cells cultured in 2AμM simvastatin NM and HCM were significantly lower than cultured in NM and HCM cells.2, the Hedgehog protein levels of prostate cancer cells cultured in 2.4μM simvastatin NM and HCM were significantly lower than that in NM and HCM, it is a significant difference between the two.3, Hedgehog signaling pathway-related protein of prostate cancer cells cultured in 2.4μM simvastatin NM and HCM, the expression Hedgehog, Smo and the Gli increased and Ptc expression decreased.4, we also observed that the 2.4μM simvastatin NM and HCM group of prostate cancer cells express Hedgehog protein in cells was significantly higher than NM and HCM groups by laser confocalConclusions:1, simvastatin inhibit prostate cancer PC-3 cells, LNCaP cells and LNCaP-AI cells in vitro.2, simvastatin inhibit Hedgehog protein in the process of extracellular secretion of prostate cancer PC-3 cells, LNCaP cells and LNCaP-AI cells.3, simvastatin inhibit prostate cancer cell proliferation correlate with the fall of extracellular secretion of Hedgehog.4, prostate cancer patients apply the lipid-lowering drugs can slow the progress of prostate cancer.
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