| Background and purpose:In recent years, the incidence of squamous cell carcinoma is going up, which is an aggressive tumor and the prognosis is very bad , but there are no good clinical treatment methods. Ursolic acid is extracted from a kind of natural plants.Many research shows that, UA can inhibit the growing of a wide variety of tumors. But the exact mechanism of anti-tumor and has not yet been fully clear, currently , the reports of UA on squamous cell carcinoma are rare. Nuclear transcription factors kappa B (NF-кB), which is a transcription regulation factor found in recent years is closely related with the tumor. NF-кB can inhibit tumor cell apoptosis by regulate many genes such as bcl-2 family, IAPs family, which are tumor antiapoptotic related gene. Studies have shown that in many human cancer, such as colon cancer, liver cancer, NF-кB signaling pathways plays an important role. This study discuss whether UA can induce the apoptosis of squamous cell carcinoma in vivo in vitro through the NF-кB pathway, In ord to provide a basis for clinical use.Methods:1. Cell culture: culture human skin squamous cell carcinoma cell line A431 cells and human skin keratinocyte cell line HaCaT cells.2.Tumor inoculation and drug delivery: A431 cells were seeded in male BALB / c nude mice left armpit, 10 days after inoculation administered, by intraperitoneal injection for UA and 5-Fu, every other day injection of 10 Times.3. Tumor identification: tumor tissue is identified by slices HE.4. Drug Safety Observation: To observe the physical condition of mice after administration, including diet, urine and mental status, and liver, lienal pathological cell necrosis observation.5. UA tumor suppressor role observation: one measuring tumor size every other day a (mm), and perpendicular to the short diameter b (mm), and using the formula V (mm3) = (a×b2)×1 / 2 calculation of the volume of tumor after 20 strip Weighing.. Inhibition rate of the control group, the average tumor weight (%) = (the average tumor weight - experimental group) / control group mean tumor weight×100%.6. The role of UA on the activity of the skin squamous cell carcinoma cell proliferation: the MTT assay and cell proliferation activity screening appropriate concentration.7. The role of UA on apoptosis rate of squamous cell carcinoma of the skin: use flow cytometry and Hoechst staining to detect cell apoptosis level.8. The effect of UA on NF-кB signaling pathway protein expression: cell protein were extracted different times after treatment, with Western blot, pαI,кBBcl -2, c-IAP-2 protein levels and IкBαResult1. Inoculated tumor identification: The identification of the pathology of tissue to the skin squamous cell carcinoma.2. Drug Safety: physical side effects of UA group is less than 5-FU group.3. Tumor suppressor role of UA: UA could significantly inhibit the growth of nude mouse skin squamous cell carcinoma, and concentration dependence.4. The role of UA and 5-FU with Different concentrations on the skin squamous cell carcinoma cell proliferation activity: The A431 cell proliferation was significantly inhibited by UA on in a time - dose dependent manner. The role of UA on the proliferation of HaCaT cells is much less than 5-FU.5. The effect of UA on apoptosis rate of squamous cell carcinoma of the skin: UA could induce the apoptosis of A431 cell in a concentration-dependent, but apoptosis of HaCaT cells was lower than 5-FU.6. The expression of NF-кB signaling pathway protein: The level of pIкBα, Bcl-2, c-IAP-2 expression decreased, The level of IкBαprotein gradually increased.ConclusionUA has obvious inhibition to transplant tumor of nude mice for skin cancer, concentration dependence. UA has obvious inhibition to A431 cells. A431 UA can obviously inducing apoptosis, and have more specific killer. UA may seem by blocking NF - and b-channel Bcl - 2 family and IAPS family etc antiapoptotic gene expression, thereby reducing the skin cells induced apoptosis scale.
|
PDF Full Text Request