中华妇产科杂志
中華婦產科雜誌
중화부산과잡지
CHINESE JOUNAL OF OBSTETRICS AND GYNECOLOGY
2008年
6期
445-450
,共6页
绒毛膜癌%肿瘤细胞,培养的%滋养层%HLA抗原%组织相容性抗原I类%细胞增殖
絨毛膜癌%腫瘤細胞,培養的%滋養層%HLA抗原%組織相容性抗原I類%細胞增殖
융모막암%종류세포,배양적%자양층%HLA항원%조직상용성항원I류%세포증식
Choriocarcinoma%Tumor eeHs,cultured%Trophoblasts%HLA antigens%Histocompatibility antigens class I: Cell proliferation
目的 探讨人类白细胞抗原G(HLA-G)基因表达水平下降对具有滋养细胞特点的绒毛膜癌(绒癌)细胞株JEG-3细胞增殖和侵袭能力的影响,了解HLA-G基因在子痫前期发生、发展中的作用.方法 采用高表达HLA-G的JEG-3细胞进行体外培养,将培养好的细胞分为实验组(转染HLA.G小分子干扰RNA)、阴性对照组(转染阴性对照小分子干扰RNA)和空白对照组(不进行细胞转染,仅转染脂质体)3组进行细胞转染.应用RT-PCR技术和蛋白印迹法检测转染后各组JEG-3细胞中HLA-G mRNA和蛋白的表达水平;四甲基偶氮唑蓝(MTT)比色法、流式细胞技术、穿膜小室侵袭实验分别检测转染后各组JEG-3细胞增殖能力、细胞周期、凋亡抑制率和侵袭能力的变化.结果 (1)转染后JEG-3细胞中HLA-G mRNA和蛋白的表达水平,实验组分别为0.0013±0.0014、0.0163±0.O007.分别与空白对照组(分别为0.1923 ±0.0384、0.2184 ±0.0153)比较,差异均有统计学意义(P<O.05);阴性对照组分别为(0.1606 ±0.0133、0.2020 ±0.0132),分别与空白对照组比较,差异均无统计学意义(P>0.05).(2)转染48 h后JEG-3细胞的增殖能力,实验组的积分吸光度(伪)值为0.44±0.04,与空白对照组(O.75 ±0.13)比较,差异有统计学意义(P<0.01);阴性对照组为0.69 ±0.10,与空白对照组比较,差异无统计学意义(P>0.05).(3)转染72 h后的细胞周期比例,实验组G2/M期细胞比例为(10.9 ±2.2)%,低于空白对照组[(15.4 ±1.9)%];S期细胞比例为(58.6±0.8)%,高于空白对照组[(52.9 ±2.3)%].两组分别比较,差异均有统计学意义(P<0.05).(4)转染72 h后的细胞凋亡率,实验组为(14.5±2.7)%,高于阴性对照组[(5.3 ±1.1)%]和空白对照组[(4.7 ±0.6)%],差异均有统计学意义(P<0.01).(5)转染72 h后穿膜小室中的穿膜细胞数,实验组为(121±12)个,低于空白对照组[(452 ±17)个],差异有统计学意义(P<0.01).结论 HLA-G基因表达水平的下降可以影响滋养细胞的增殖、凋亡、侵袭和细胞周期.HLA-G基因可能通过调节滋养细胞增殖、侵袭等过程参与子痫前期的发生.
目的 探討人類白細胞抗原G(HLA-G)基因錶達水平下降對具有滋養細胞特點的絨毛膜癌(絨癌)細胞株JEG-3細胞增殖和侵襲能力的影響,瞭解HLA-G基因在子癇前期髮生、髮展中的作用.方法 採用高錶達HLA-G的JEG-3細胞進行體外培養,將培養好的細胞分為實驗組(轉染HLA.G小分子榦擾RNA)、陰性對照組(轉染陰性對照小分子榦擾RNA)和空白對照組(不進行細胞轉染,僅轉染脂質體)3組進行細胞轉染.應用RT-PCR技術和蛋白印跡法檢測轉染後各組JEG-3細胞中HLA-G mRNA和蛋白的錶達水平;四甲基偶氮唑藍(MTT)比色法、流式細胞技術、穿膜小室侵襲實驗分彆檢測轉染後各組JEG-3細胞增殖能力、細胞週期、凋亡抑製率和侵襲能力的變化.結果 (1)轉染後JEG-3細胞中HLA-G mRNA和蛋白的錶達水平,實驗組分彆為0.0013±0.0014、0.0163±0.O007.分彆與空白對照組(分彆為0.1923 ±0.0384、0.2184 ±0.0153)比較,差異均有統計學意義(P<O.05);陰性對照組分彆為(0.1606 ±0.0133、0.2020 ±0.0132),分彆與空白對照組比較,差異均無統計學意義(P>0.05).(2)轉染48 h後JEG-3細胞的增殖能力,實驗組的積分吸光度(偽)值為0.44±0.04,與空白對照組(O.75 ±0.13)比較,差異有統計學意義(P<0.01);陰性對照組為0.69 ±0.10,與空白對照組比較,差異無統計學意義(P>0.05).(3)轉染72 h後的細胞週期比例,實驗組G2/M期細胞比例為(10.9 ±2.2)%,低于空白對照組[(15.4 ±1.9)%];S期細胞比例為(58.6±0.8)%,高于空白對照組[(52.9 ±2.3)%].兩組分彆比較,差異均有統計學意義(P<0.05).(4)轉染72 h後的細胞凋亡率,實驗組為(14.5±2.7)%,高于陰性對照組[(5.3 ±1.1)%]和空白對照組[(4.7 ±0.6)%],差異均有統計學意義(P<0.01).(5)轉染72 h後穿膜小室中的穿膜細胞數,實驗組為(121±12)箇,低于空白對照組[(452 ±17)箇],差異有統計學意義(P<0.01).結論 HLA-G基因錶達水平的下降可以影響滋養細胞的增殖、凋亡、侵襲和細胞週期.HLA-G基因可能通過調節滋養細胞增殖、侵襲等過程參與子癇前期的髮生.
목적 탐토인류백세포항원G(HLA-G)기인표체수평하강대구유자양세포특점적융모막암(융암)세포주JEG-3세포증식화침습능력적영향,료해HLA-G기인재자간전기발생、발전중적작용.방법 채용고표체HLA-G적JEG-3세포진행체외배양,장배양호적세포분위실험조(전염HLA.G소분자간우RNA)、음성대조조(전염음성대조소분자간우RNA)화공백대조조(불진행세포전염,부전염지질체)3조진행세포전염.응용RT-PCR기술화단백인적법검측전염후각조JEG-3세포중HLA-G mRNA화단백적표체수평;사갑기우담서람(MTT)비색법、류식세포기술、천막소실침습실험분별검측전염후각조JEG-3세포증식능력、세포주기、조망억제솔화침습능력적변화.결과 (1)전염후JEG-3세포중HLA-G mRNA화단백적표체수평,실험조분별위0.0013±0.0014、0.0163±0.O007.분별여공백대조조(분별위0.1923 ±0.0384、0.2184 ±0.0153)비교,차이균유통계학의의(P<O.05);음성대조조분별위(0.1606 ±0.0133、0.2020 ±0.0132),분별여공백대조조비교,차이균무통계학의의(P>0.05).(2)전염48 h후JEG-3세포적증식능력,실험조적적분흡광도(위)치위0.44±0.04,여공백대조조(O.75 ±0.13)비교,차이유통계학의의(P<0.01);음성대조조위0.69 ±0.10,여공백대조조비교,차이무통계학의의(P>0.05).(3)전염72 h후적세포주기비례,실험조G2/M기세포비례위(10.9 ±2.2)%,저우공백대조조[(15.4 ±1.9)%];S기세포비례위(58.6±0.8)%,고우공백대조조[(52.9 ±2.3)%].량조분별비교,차이균유통계학의의(P<0.05).(4)전염72 h후적세포조망솔,실험조위(14.5±2.7)%,고우음성대조조[(5.3 ±1.1)%]화공백대조조[(4.7 ±0.6)%],차이균유통계학의의(P<0.01).(5)전염72 h후천막소실중적천막세포수,실험조위(121±12)개,저우공백대조조[(452 ±17)개],차이유통계학의의(P<0.01).결론 HLA-G기인표체수평적하강가이영향자양세포적증식、조망、침습화세포주기.HLA-G기인가능통과조절자양세포증식、침습등과정삼여자간전기적발생.
Objective To investigate the effect of human leukocyte antigen-G(HLA-c)on the growth and invasion of JEG-3 cell line and the role of HLA-G in the onset and development of pre-eclampsia.Methods The experiment was composed of three groups:groups of transfection,negative control and blank control.which corresponded to groups of HLA.G siRNA transfection,negative siRNA transfection and no transfection HLA-G overexpressed choriocarcinoma cell line JEG-3 was used.The role of HLA-G in JEG-3cell monolayer was examined by RNA interference technology using HLA-G specific small interfering RNA (siRNA).Expression of HLA.G was detected by reverse transeriptase-polymerase chain reaction and western blot analysis.Changes of cell cycle,apoptosis,proliferation and invasion were respectively detected by methvl thiazolyl tetrazolium(Ma r).flow cytometry assay and transwell test.Results (1)The mRNA and protein levels of HLA.G control group and blank control group were 0.0013±0.0014.0.0163 ±0.0007 and 0.1923 ±0.0384.0.2184 ±0.0153,respectively,which were both significantly different(P<0.05);the number of negative transfcction group was 0.1606±0.0133 and 0.2020±0.0132.which had no significant difference compared with blank control group(P>0.05).(2)The integral absorbance(IA)valUCB of the HLA-G transfecfion group and blank control group were 0.44±0.04 and 0.75±0.13 respectively.which was significantly different(P<0.01);the/A value of negative control group was 0.69±0.10.which was not significantly different compared with blank group(P>0.05).(3)The ratios of G2/M and S phase cells in transfection group were(10.9±2.2)%and(58.6±0.8)%respectively,significantly different compared with the blank control group[(15.4±1.9)%and(52.9±2.3)%respectively;P<0.01].(4)The ratio of early apoptosis cells in transfection group[(14.5±2.7)%]Was significantly increased compared with neg~ive[(5.3 ±1.1)%]and blank control group[(4.7±0.6)%;P<0.01].(5)The invasion number of transfecfion group and blank control group were 121±12 and 452±17 respectively.with a significant eclampsia by regulating proliferation and invasion of trophoblast.
