中国癌症杂志
中國癌癥雜誌
중국암증잡지
CHINA ONCOLOGY
2013年
11期
868-873
,共6页
陆媛媛%张洁清%梁少凤%李力
陸媛媛%張潔清%樑少鳳%李力
륙원원%장길청%량소봉%리력
雌二醇%子宫内膜癌%AKT%增殖
雌二醇%子宮內膜癌%AKT%增殖
자이순%자궁내막암%AKT%증식
Estrogen%Endometrial cancer%AKT%Proliferation
背景与目的：目前认为子宫内膜癌的发生可能与无拮抗的雌激素长期作用有关，但雌激素如何调节细胞增殖的作用机制尚不清楚。AKT信号转导通路是细胞生存和增殖的一个重要调节信号。本研究探讨在子宫内膜癌细胞HEC-1A中，雌二醇能否通过激活AKT通路产生细胞因子，以及其对细胞增殖能力的影响。方法：蛋白质印迹法(Western blot)技术检测雌二醇作用HEC-1A细胞后AKT活化情况，以及AKT抑制剂、ER抑制剂对AKT活化的影响。荧光定量PCR及ELISA技术检测雌二醇(E2组)作用于HEC-1A细胞30 min后；或雌激素受体抑制剂(ER组)、AKT抑制剂(AKT组)分别预处理细胞1 h后再加入雌二醇作用30 min后，细胞内血管内皮生长因子受体(VEGF)、bFGF、IL-8基因mRNA表达及细胞培养上清液中蛋白表达。细胞集落形成实验、流式细胞仪检测细胞周期的变化，CFSE法检测细胞增殖能力。结果：E2组的AKT活化比值较对照组显著升高(P=0.0062)，ER组和AKT组的AKT活性比值较E2组显著降低(P=0.0060和P=0.0064)，但不能完全抑制雌二醇作用。E2组的VEGF、bFGF、IL-8 mRNA或蛋白的表达均明显高于对照组(P均<0.01)；在ER组及AKT组中VEGF、bFGF、IL-8 mRNA或蛋白的表达均明显低于E2组(P均<0.05)；雌二醇作用HEC-1A细胞后，细胞增殖数明显增多，细胞周期加快(P均<0.01)。结论：在HEC-1A细胞，雌二醇可能通过激活AKT信号通路产生VEGF、bFGF、IL-8因子，从而增强肿瘤细胞的增殖能力。
揹景與目的：目前認為子宮內膜癌的髮生可能與無拮抗的雌激素長期作用有關，但雌激素如何調節細胞增殖的作用機製尚不清楚。AKT信號轉導通路是細胞生存和增殖的一箇重要調節信號。本研究探討在子宮內膜癌細胞HEC-1A中，雌二醇能否通過激活AKT通路產生細胞因子，以及其對細胞增殖能力的影響。方法：蛋白質印跡法(Western blot)技術檢測雌二醇作用HEC-1A細胞後AKT活化情況，以及AKT抑製劑、ER抑製劑對AKT活化的影響。熒光定量PCR及ELISA技術檢測雌二醇(E2組)作用于HEC-1A細胞30 min後；或雌激素受體抑製劑(ER組)、AKT抑製劑(AKT組)分彆預處理細胞1 h後再加入雌二醇作用30 min後，細胞內血管內皮生長因子受體(VEGF)、bFGF、IL-8基因mRNA錶達及細胞培養上清液中蛋白錶達。細胞集落形成實驗、流式細胞儀檢測細胞週期的變化，CFSE法檢測細胞增殖能力。結果：E2組的AKT活化比值較對照組顯著升高(P=0.0062)，ER組和AKT組的AKT活性比值較E2組顯著降低(P=0.0060和P=0.0064)，但不能完全抑製雌二醇作用。E2組的VEGF、bFGF、IL-8 mRNA或蛋白的錶達均明顯高于對照組(P均<0.01)；在ER組及AKT組中VEGF、bFGF、IL-8 mRNA或蛋白的錶達均明顯低于E2組(P均<0.05)；雌二醇作用HEC-1A細胞後，細胞增殖數明顯增多，細胞週期加快(P均<0.01)。結論：在HEC-1A細胞，雌二醇可能通過激活AKT信號通路產生VEGF、bFGF、IL-8因子，從而增彊腫瘤細胞的增殖能力。
배경여목적：목전인위자궁내막암적발생가능여무길항적자격소장기작용유관，단자격소여하조절세포증식적작용궤제상불청초。AKT신호전도통로시세포생존화증식적일개중요조절신호。본연구탐토재자궁내막암세포HEC-1A중，자이순능부통과격활AKT통로산생세포인자，이급기대세포증식능력적영향。방법：단백질인적법(Western blot)기술검측자이순작용HEC-1A세포후AKT활화정황，이급AKT억제제、ER억제제대AKT활화적영향。형광정량PCR급ELISA기술검측자이순(E2조)작용우HEC-1A세포30 min후；혹자격소수체억제제(ER조)、AKT억제제(AKT조)분별예처리세포1 h후재가입자이순작용30 min후，세포내혈관내피생장인자수체(VEGF)、bFGF、IL-8기인mRNA표체급세포배양상청액중단백표체。세포집락형성실험、류식세포의검측세포주기적변화，CFSE법검측세포증식능력。결과：E2조적AKT활화비치교대조조현저승고(P=0.0062)，ER조화AKT조적AKT활성비치교E2조현저강저(P=0.0060화P=0.0064)，단불능완전억제자이순작용。E2조적VEGF、bFGF、IL-8 mRNA혹단백적표체균명현고우대조조(P균<0.01)；재ER조급AKT조중VEGF、bFGF、IL-8 mRNA혹단백적표체균명현저우E2조(P균<0.05)；자이순작용HEC-1A세포후，세포증식수명현증다，세포주기가쾌(P균<0.01)。결론：재HEC-1A세포，자이순가능통과격활AKT신호통로산생VEGF、bFGF、IL-8인자，종이증강종류세포적증식능력。
Background and purpose:The occurrence of endometrial cancer may be related to the persistent stimulus of endogenous and exogenous estrogen without progesterone antagonist. But how does estrogen regulate cell proliferation is still unknown. AKT pathway is the most important signal transduction way to mediate proliferation in the cells. The main aim was to study whether estradiol induces the expression of VEGF, bFGF and IL-8 in the endometrial cancer HEC-1A cells by activating AKT, and its effect on proliferation. Methods:Western blot was used to detect the expression of AKT protein in HEC-1A cells after estradiol stimulation, AKT inhibitor or ER inhibitor stimulation followed by estradiol. Real-time PCR and ELISA were used to detect the gene and protein expression of VEGF, bFGF and IL-8 in different inhibitors. Cell colony formation assay, lfow cytometry and CFSE assay were used to examine the proliferation in HEC-1A cells. Results:The expression of p-AKT protein in HEC-1A cells after stimulation with estradiol was markedly higher than that in the control group (P=0.006 2);the expression of p-AKT protein in AKT inhibitor group and ER inhibitor group were signiifcantly decreased than that in estradiol group (P=0.006 0, P=0.006 4). qPCR and ELISA showed the mRNA and protein expression of VEGF, bFGF, IL-8 in estradiol group were signiifcantly increased than that in control group (P<0.05);The expressions of VEGF, bFGF, IL-8 in AKT inhibitor group and ER inhibitor group were signiifcantly decreased than that in estradiol group (P<0.01). The abilities of proliferation and cell cycle were signiifcantly increased in HEC-1A cells after estradiol stimulation. Conclusion:Estrogen induces the production of VEGF, bFGF and IL-8 through activating AKT signal pathway.
