黄芩素对宫颈癌Hela细胞增殖的抑制作用及机制探讨
황금소대궁경암Hela세포증식적억제작용급궤제탐토
Inhibitory effect of baicalein on proliferation of cervical carcinoma Hela cells and its mechanism
  • 万方数据
    山东医药 산동의약
    SHANDONG MEDICAL JOURNAL
    2014年 27期 4-7 ,共4页

    卢科莲%喻小兰%夏纪毅%毛熙光

    로과련%유소란%하기의%모희광

    黄芩素%宫颈癌%宫颈肿瘤%Hela细胞%基质金属蛋白酶%大鼠肉瘤蛋白%细胞周期素D1%整合素β1 黃芩素%宮頸癌%宮頸腫瘤%Hela細胞%基質金屬蛋白酶%大鼠肉瘤蛋白%細胞週期素D1%整閤素β1
    황금소%궁경암%궁경종류%Hela세포%기질금속단백매%대서육류단백%세포주기소D1%정합소β1
    baicalein%cervical carcinoma%cervical neoplasms%Hela cells%matrix metalloproteinases%murine sarcoma protein%Cyclin D1%integrin β1
    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。
    목적：관찰황금소대궁경암Hela세포증식적억제작용，병탐토기가능궤제。방법체외배양적궁경암Hela세포분위3조，A조가입100μmol/L황금소， B조가입200μmol/L황금소， C조가입불함황금소적무혈청DMEM고당배양기，처리24 h。상차현미경관찰세포형태급수량，류식세포기술검측세포주기，명효매보법검측세포배양기중기질금속단백매（ MMP）-2화MMP-9활성，RT-PCR법검측MMP-2、MMP-9、대서육류단백（ Ras）、세포주기소 D1（ CyclinD1）、정합소β1（ Integrinβ1） mRNA， Western blotting 법검측 MMP-2、MMP-9、Ras、CyclinD1、Integrinβ1단백。결과여C조상비교，A、B조세포수량감소，세포형태종불규칙류원형전변위다각사형。 A、B、C조S기세포백분비분별위24．58％、19．41％、34．05％，G1기세포백분비분별위72．67％、79．17％、64．88％，A、B조여C조비교，P균＜0．05。 A、B、C조배양기중MMP-2활성분별위32．651±0．571、28．987±1．033、36．127±1．184， MMP-9활성분별위33．217±0．660、29．277±0．868、39．613±0．318，A、B조여C조비교，P균＜0．05。 A조MMP-2、MMP-9、Ras、CyclinD1、Integrinβ1 mRNA표체수평분별위1．012±0．045、1．009±0．030、0．988±0．036、1．013±0．034、0．983±0．036，B조분별위0．849±0．026、0．914±0．027、0．626±0．029、0．806±0．037、0．799±0．037，C조분별위1．199±0．029、1．087±0．024、1．413±0．026、1．203±0．029、1．224±0．025，A、B조여C조비교，P균＜0．05。 A조MMP-2、MMP-9、Ras、CyclinD1、Integrinβ1단백표체수평분별위0．693±0．025、0．886±0．032、1．127±0．028、1．061±0．028、1．159±0．031，B조분별위0．326±0．022、0．754±0．024、0．782±0．183、0．321±0．027、0．481±0．028，C조분별위1．914±0031、1．487±0．016、1．323±0．303、1．816±0．025、1．467±0．036，A、B조여C조비교，P균＜0．05。결론황금소가억제궁경암Hela세포증식，기궤제가능위장세포조체우G1기，강저세포배양기중MMP-2、MMP-9활성，하조세포MMP-2、MMP-9、Ras、CyclinD1、Integrinβ1 mRNA급단백표체。
    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<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<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 statistical difference was found between groups A , B and C (all P<0.05);the protein expression of MMP-2, MMP-9, Ras, Cyclin D1 and integrinβ1 in the group A was 0.693 ±0.025, 0.886 ±0.032, 1.127 ±0.028, 1.061 ±0.028 and 1.159 ±0.031, in the group B was 0.326 ±0.022, 0.754 ±0.024, 0.782 ±0.183, 0.321 ±0.027 and 0.481 ±0.028, and in the group C was 1.914 ±0031, 1.487 ±0.016, 1.323 ±0.303, 1.816 ±0.025 and 1.467 ±0.036;and statistical difference was found between groups A , B and C (all P<0.05).Conclusion Baicalein inhibits the proliferation of cervical cancer Hela cells by arresting the cell in the G 1 phase, decreasing the activities of MMP-2 and MMP-9, and down-regulating the mRNA and protein expression of MMP-2, MMP-9, Ras, Cyclin D1 and integrin β1.
    원문보기 원문다운로드 사이트로이동
저자의 최근 논문