摘要
目的观察黄芩素对宫颈癌Hela细胞增殖的抑制作用,并探讨其可能机制。方法体外培养的宫颈癌Hela细胞分为3组,A组加入100μmol/L黄芩素,B组加入200μmol/L黄芩素,C组加入不含黄芩素的无血清DMEM高糖培养基,处理24 h。相差显微镜观察细胞形态及数量,流式细胞技术检测细胞周期,明胶酶谱法检测细胞培养基中基质金属蛋白酶(MMP)-2和MMP-9活性,RT-PCR法检测MMP-2、MMP-9、大鼠肉瘤蛋白(Ras)、细胞周期素D1(CyclinD1)、整合素β1(Integrinβ1)mRNA,Western blotting法检测MMP-2、MMP-9、Ras、CyclinD1、Integrinβ1蛋白。结果与C组相比较,A、B组细胞数量减少,细胞形态从不规则类圆形转变为多角梭形。A、B、C组S期细胞百分比分别为24.58%、19.41%、34.05%,G1期细胞百分比分别为72.67%、79.17%、64.88%,A、B组与C组比较,P均<0.05。A、B、C组培养基中MMP-2活性分别为32.651±0.571、28.987±1.033、36.127±1.184,MMP-9活性分别为33.217±0.660、29.277±0.868、39.613±0.318,A、B组与C组比较,P均<0.05。A组MMP-2、MMP-9、Ras、CyclinD1、Integrinβ1 mRNA表达水平分别为1.012±0.045、1.009±0.030、0.988±0.036、1.013±0.034、0.983±0.036,B组分别为0.849±0.026、0.914±0.027、0.626±0.029、0.806±0.037、0.799±0.037,C组分别为1.199±0.029、1.087±0.024、1.413±0.026、1.203±0.029、1.224±0.025,A、B组与C组比较,P均<0.05。A组MMP-2、MMP-9、Ras、CyclinD1、Integrinβ1蛋白表达水平分别为0.693±0.025、0.886±0.032、1.127±0.028、1.061±0.028、1.159±0.031,B组分别为0.326±0.022、0.754±0.024、0.782±0.183、0.321±0.027、0.481±0.028,C组分别为1.914±0031、1.487±0.016、1.323±0.303、1.816±0.025、1.467±0.036,A、B组与C组比较,P均<0.05。结论黄芩素可抑制宫颈癌Hela细胞增殖,其机制可能为将细胞阻滞于G1期,降低细胞培养基中MMP-2、MMP-9活性,下调细胞MMP-2、MMP-9、Ras、CyclinD1、Integrinβ1 mRNA及蛋白表达。
Objective To observe the inhibitory effect of baicalein on the proliferation of cervical carcinoma Hela cells and to investigate its possible mechanism .Methods Hela cells cultured in vitro were randomly divided into 3 groups：group A which was added 100μmol/L baicalein, group B which was added 200μmol/L baicalein and group C which was added se-rum-free DMEM high-glucose medium without baicalein , and they were all treated for 24 hours.The morphology and quantity of Hela cells were observed under phase-contrast microscope , the cell cycle of each group was detected by flow cytometry , the activities of matrix metalloproteinases （ MMP-2 and MMP-9 ） were detected by gelatin zymography , the mRNA expression changes of MMP-2, MMP-9, murine sarcoma protein （Ras）, Cyclin D1 and integrinβ1 were measured by RT-PCR, and pro-tein expression changes of MMP-2, MMP-9, Ras, Cyclin D1 and integrinβ1 were measured by Western blotting .Results Compared with group C , the cell quantities of groups A and B were decreased , and the cell morphology changed from irregular similar circular to multi-angle spindle;the cell percentages in the S phase of groups A , B and C were 24.58%, 19.41%and 34.05%;and the cell percentages in the G 1 phase were 72.67%, 79.17%and 64.88%, respectively;and statistical differ-ence was found between groups A , B and C （all P＆lt;0.05）;the activities of MMP-2 in groups A, B and C were 32.651 ± 0.571, 28.987 ±1.033 and 36.127 ±1.184;the activities of MMP-9 in groups A, B and C were 33.217 ±0.660, 29.277 ± 0.868 and 39.613 ±0.318;and statistical difference was found between groups A , B and C （ all P＆lt;0.05）;the mRNA ex-pression of MMP-2, MMP-9, Ras, Cyclin D1 and integrin β1 in the group A was 1.012 ±0.045, 1.009 ±0.030, 0.988 ± 0.036, 1.013 ±0.034 and 0.983 ±0.036;it was 0.849 ±0.026, 0.914 ±0.027, 0.626 ±0.029, 0.806 ±0.037 and 0.799 ±0.037 in the group B;it was 1.199 ±0.029, 1.087 ±0.024, 1.413 ±0.026, 1.203 ±0.029 and 1.224 ±0.025 in the group C, and statisti
出处
《山东医药》
CAS
2014年第27期4-7,共4页
Shandong Medical Journal
基金
四川省卫生厅基金资助项目(12193)
四川省科学技术厅课题基金(09032)
