【摘要】：目的觀察中藥苦參堿(Matrine)對宮頸癌SiHa細(xì)胞增殖的抑制作用以及對JAK-STAT (Janus Kinase/Signal Transducers and Actiators of Transcription)信號(hào)轉(zhuǎn)導(dǎo)通路的影響。 方法1體外培養(yǎng)宮頸癌SiHa細(xì)胞,觀察細(xì)胞的生長狀態(tài),取處于對數(shù)生長期的細(xì)胞用于試驗(yàn)。2試驗(yàn)分組：(1)實(shí)驗(yàn)組：用不同濃度的苦參堿處理宮頸癌SiHa細(xì)胞,使其終濃度分別為0.25mg/ml、0.5mg/ml、1.0mg/ml、2.0mg/ml、4.0mg/ml;(2)對照組：只加入細(xì)胞懸液,未經(jīng)藥物處理；(3)空白對照組：不接種細(xì)胞,僅加入細(xì)胞培養(yǎng)液。3應(yīng)用MTT法檢測不同濃度苦參堿(0.25mg/ml、0.5mg/ml、1.0mg/ml、2.0mg/ml、4.0mg/ml)作用于宮頸癌SiHa細(xì)胞24h、48h及72h后細(xì)胞的增殖情況。4采用蛋白印跡法(Western blotting)法檢測不同濃度苦參堿(0.5mg/ml、1.0mg/ml、2.0mg/ml)作用于宮頸癌SiHa細(xì)胞48h后,SiHa細(xì)胞中JAK1、JAK2、STAT3蛋白的表達(dá)水平。 結(jié)果1苦參堿能夠抑制體外培養(yǎng)的宮頸癌SiHa細(xì)胞的增殖,該抑制作用呈時(shí)間-劑量依賴性。各組不同作用時(shí)間比較,苦參堿對宮頸癌SiHa細(xì)胞增殖的抑制率差異顯著(P0.05、0.01)。當(dāng)苦參堿的加入濃度在0.25mg/ml～4.0mg/ml的范圍時(shí),苦參堿的濃度越高,其對宮頸癌SiHa細(xì)胞的增殖抑制效應(yīng)越明顯；2不同濃度的苦參堿(0.5mg/ml、1.0mg/ml、2.0mg/ml)作用于宮頸癌SiHa細(xì)胞48h后,各實(shí)驗(yàn)組中JAK1、JAK2和STAT3蛋白的表達(dá)水平降低(P0.01),且有劑量依賴性。 結(jié)論苦參堿對宮頸癌SiHa細(xì)胞有抑制作用。其作用機(jī)制可能是通過降低宮頸癌細(xì)胞中JAK-STAT信號(hào)轉(zhuǎn)導(dǎo)通路的活性,從而抑制宮頸癌細(xì)胞的增殖,有望成為治療宮頸癌以及抑制其侵襲轉(zhuǎn)移的一種新的策略。
[Abstract]:Aim to observe the inhibitory effect of matrine (Matrine) on the proliferation of cervical cancer SiHa cells and the effect of JAK-STAT (Janus Kinase/Signal Transducers and Actiators of Transcription) signal transduction pathway). Methods (1) Cervical carcinoma SiHa cells were cultured in vitro, the growth state of the cells was observed, and the cells in logarithmic growth phase were used in the experiment. 2 the experimental groups were as follows: (1) the experimental group: the cervical cancer SiHa cells were treated with different concentrations of matrine. The final concentration is 0.25 mg / ml, 0.5 mg / ml, 1.0 mg / ml, 2.0 mg / ml, 4.0 mg / ml, respectively; (2) Control group: only cell suspension was added without drug treatment; (3) blank control group: no cells were inoculated, only cell culture medium was added. (3) different concentrations of matrine (0.25 mg / ml, 0.5 mg / ml, 1.0 mg / ml, 2.0 mg / ml) were detected by MTT method. (4) the proliferation of cervical cancer SiHa cells was detected by Western blot (Western blotting) assay after 24 h and 72 h treatment with 4.0mg/ml (0.5 mg 路ml ~ (-1) 路ml ~ (-1) ~ (-1) mg 路ml ~ (-1) of matrine (0.5 mg 路ml ~ (-1) 路ml ~ (-1), respectively. The expression of JAK1,JAK2,STAT3 protein in SiHa cells of cervical cancer treated with 2.0mg/ml for 48 h. Results 1 Matrine could inhibit the proliferation of cervical cancer SiHa cells in a time-dose-dependent manner. The inhibitory rate of matrine on the proliferation of cervical cancer SiHa cells was significantly different (P0.05 / 0. 01). When the concentration of matrine was in the range of 0.25mg/ml~4.0mg/ml, the higher the concentration of matrine was, the more obvious the inhibitory effect of matrine on the proliferation of cervical cancer SiHa cells was. 2 different concentrations of matrine (0.5 mg / ml, 1.0 mg / ml, 2.0 mg / ml) were treated with cervical cancer SiHa cells for 48 h, the expression levels of JAK1,JAK2 and STAT3 protein in each experimental group were decreased (P0.01) in a dose-dependent manner. Conclusion Matrine can inhibit SiHa cells of cervical cancer. Its mechanism may be to inhibit the proliferation of cervical cancer cells by reducing the activity of JAK-STAT signal transduction pathway in cervical cancer cells, which may become a new strategy for the treatment of cervical cancer and its invasion and metastasis.
【學(xué)位授予單位】：蘭州大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R737.33

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