中华肝脏病杂志
中華肝髒病雜誌
중화간장병잡지
CHINESE JOURNAL OF HEPATOLOGY
2012年
12期
925-929
,共5页
徐玫芳%臧盛兵%刘景丰%高凌云%高美钦%杨映红%黄爱民
徐玫芳%臧盛兵%劉景豐%高凌雲%高美欽%楊映紅%黃愛民
서매방%장성병%류경봉%고릉운%고미흠%양영홍%황애민
癌,肝细胞%细胞增殖%基因,肿瘤抑制
癌,肝細胞%細胞增殖%基因,腫瘤抑製
암,간세포%세포증식%기인,종류억제
Carcinoma,hepatocellular%Cell proliferation%Genes,tumor suppressor
目的 观察KiSS-1基因表达对肝癌细胞体外增殖、黏附及侵袭能力的影响,为进一步探讨其抗肝细胞癌侵袭转移的机制奠定基础.方法 培养具有高转移潜能的人肝癌细胞株MHCC97-H,瞬时转染KiSS-1基因的细胞为实验组,转染空载体pcDNA3.1/HisC的细胞为空白对照组,未转染细胞为阴性对照组,采用流式细胞术与四甲基偶氮唑盐法、基质黏附实验、Transwell体外侵袭和趋化运动实验检测KiSS-1表达对MHCC97-H细胞体外增殖、黏附、侵袭和运动能力的影响.应用SPSS13.0统计软件包进行统计分析,组间差异以两样本t检验分析. 结果 转染组、空载体组和未转染组与Matrigel基质的黏附能力(A值)分别为0.257±0.029、0.374±0.016和0.394±0.031,转染组明显低于空载体组(t=-7.90345,P＜0.01)和未转染组(t=-7.22752,P＜0.01);与Fibronectin基质的黏附能力(A值)分别为0.292±0.004、0.394±0.010和0.412±0.023,转染组明显低于空载体组(t=-20.93138,P＜0.01)和未转染组(t=-11.31371,P＜0.01)；趋化运动至Transwell小室下室表面的细胞数分别为65.80±1.92、93.80±2.28和96.40±2.07,转染组明显低于空载体组(t=-30.11750,P＜ 0.01)和未转染组(t=-24.19142,P＜0.01);侵袭至Transwell小室下室表面的细胞数为42.40±1.14、66.00±1.58和67.80±1.92,转染组明显低于空载体组(t=-27.0711,P＜0.01)和未转染组(t=-25.4,P＜0.01).而转染组、空载体组和未转染组的细胞增殖能力(A值)分别为0.644±0.027、0.669±0.022和0.678±0.027,转染组略低于空载体组(t=-1.60371,P＞0.05)和未转染组(t=-1.97828,P＞0.05);同时,转染组较空载体组和未转染组未出现明显的G1期阻滞和S期阻滞,也未出现明显的凋亡峰.结论 KiSS-1表达虽不影响肝癌细胞的体外增殖能力,但可抑制其黏附、侵袭和运动能力,提示KiSS-1可作为一个候选的肝细胞癌转移抑制基因,可望成为肝癌侵袭转移治疗的新靶点.
目的 觀察KiSS-1基因錶達對肝癌細胞體外增殖、黏附及侵襲能力的影響,為進一步探討其抗肝細胞癌侵襲轉移的機製奠定基礎.方法 培養具有高轉移潛能的人肝癌細胞株MHCC97-H,瞬時轉染KiSS-1基因的細胞為實驗組,轉染空載體pcDNA3.1/HisC的細胞為空白對照組,未轉染細胞為陰性對照組,採用流式細胞術與四甲基偶氮唑鹽法、基質黏附實驗、Transwell體外侵襲和趨化運動實驗檢測KiSS-1錶達對MHCC97-H細胞體外增殖、黏附、侵襲和運動能力的影響.應用SPSS13.0統計軟件包進行統計分析,組間差異以兩樣本t檢驗分析. 結果 轉染組、空載體組和未轉染組與Matrigel基質的黏附能力(A值)分彆為0.257±0.029、0.374±0.016和0.394±0.031,轉染組明顯低于空載體組(t=-7.90345,P＜0.01)和未轉染組(t=-7.22752,P＜0.01);與Fibronectin基質的黏附能力(A值)分彆為0.292±0.004、0.394±0.010和0.412±0.023,轉染組明顯低于空載體組(t=-20.93138,P＜0.01)和未轉染組(t=-11.31371,P＜0.01)；趨化運動至Transwell小室下室錶麵的細胞數分彆為65.80±1.92、93.80±2.28和96.40±2.07,轉染組明顯低于空載體組(t=-30.11750,P＜ 0.01)和未轉染組(t=-24.19142,P＜0.01);侵襲至Transwell小室下室錶麵的細胞數為42.40±1.14、66.00±1.58和67.80±1.92,轉染組明顯低于空載體組(t=-27.0711,P＜0.01)和未轉染組(t=-25.4,P＜0.01).而轉染組、空載體組和未轉染組的細胞增殖能力(A值)分彆為0.644±0.027、0.669±0.022和0.678±0.027,轉染組略低于空載體組(t=-1.60371,P＞0.05)和未轉染組(t=-1.97828,P＞0.05);同時,轉染組較空載體組和未轉染組未齣現明顯的G1期阻滯和S期阻滯,也未齣現明顯的凋亡峰.結論 KiSS-1錶達雖不影響肝癌細胞的體外增殖能力,但可抑製其黏附、侵襲和運動能力,提示KiSS-1可作為一箇候選的肝細胞癌轉移抑製基因,可望成為肝癌侵襲轉移治療的新靶點.
목적 관찰KiSS-1기인표체대간암세포체외증식、점부급침습능력적영향,위진일보탐토기항간세포암침습전이적궤제전정기출.방법 배양구유고전이잠능적인간암세포주MHCC97-H,순시전염KiSS-1기인적세포위실험조,전염공재체pcDNA3.1/HisC적세포위공백대조조,미전염세포위음성대조조,채용류식세포술여사갑기우담서염법、기질점부실험、Transwell체외침습화추화운동실험검측KiSS-1표체대MHCC97-H세포체외증식、점부、침습화운동능력적영향.응용SPSS13.0통계연건포진행통계분석,조간차이이량양본t검험분석. 결과 전염조、공재체조화미전염조여Matrigel기질적점부능력(A치)분별위0.257±0.029、0.374±0.016화0.394±0.031,전염조명현저우공재체조(t=-7.90345,P＜0.01)화미전염조(t=-7.22752,P＜0.01);여Fibronectin기질적점부능력(A치)분별위0.292±0.004、0.394±0.010화0.412±0.023,전염조명현저우공재체조(t=-20.93138,P＜0.01)화미전염조(t=-11.31371,P＜0.01)；추화운동지Transwell소실하실표면적세포수분별위65.80±1.92、93.80±2.28화96.40±2.07,전염조명현저우공재체조(t=-30.11750,P＜ 0.01)화미전염조(t=-24.19142,P＜0.01);침습지Transwell소실하실표면적세포수위42.40±1.14、66.00±1.58화67.80±1.92,전염조명현저우공재체조(t=-27.0711,P＜0.01)화미전염조(t=-25.4,P＜0.01).이전염조、공재체조화미전염조적세포증식능력(A치)분별위0.644±0.027、0.669±0.022화0.678±0.027,전염조략저우공재체조(t=-1.60371,P＞0.05)화미전염조(t=-1.97828,P＞0.05);동시,전염조교공재체조화미전염조미출현명현적G1기조체화S기조체,야미출현명현적조망봉.결론 KiSS-1표체수불영향간암세포적체외증식능력,단가억제기점부、침습화운동능력,제시KiSS-1가작위일개후선적간세포암전이억제기인,가망성위간암침습전이치료적신파점.
Objective To investigate the impact of expression ofkisspeptin-1 (KiSS-1) metastasissuppressor gene on the proliferative,adhesive and invasive abilities of human hepatocellular carcinoma (HCC) using an in vitro cell system.Methods The highly metastatic human hepatoma cell line MHCC97-H was transiently transfected with the pcDNA3.1/HisC vector expressing the KiSS-1 gene (experimental group) or the vector without the KisS-1 gene (blank control group).Untransfected cells served as the negative control group.Proliferative abilities of the three groups were assessed by flow cytometry and MTT assay.Adhesive abilities were assessed by MTT assays using matrigel and fibronectin.Invasive abilities and cell motility were assessed by chemoinvasion chamber assay using reconstituted matrigel and migration chamber assay using polycarbonate filters,respectively.Results The experimental group showed significantly lower adhesion capacity to matrigel (0.257 ± 0.029) than either the blank control group (0.374 ± 0.016; t=-7.90345,P＜ 0.01) or the negative control group (0.394 ± 0.031; t=-7.22752,P＜ 0.01).Similarly,the experimental group showed significantly lower adhesion capacity to fibronectin (0.292 ± 0.004) than either the blank control group (0.394 ± 0.010; t =-20.93138,P＜ 0.01) or the negative control group (0.412 ± 0.023; t=-11.31371,P ＜ 0.01).The experimental group also showed significantly lower numbers of cells with invasive capacity (42.40 ± 1.14) than either the blank control group (66 ± 1.58; t =-27.0711,P ＜ 0.01) or the negative control group (67.80 ± 1.92;t =-25.4,P ＜ 0.01).Similarly,the experimental group showed significantly lower numbers of cells with chemotactic movement (65.80 ± 1.92) than either the blank control group (93.80 ± 2.28; t =-30.11750,P ＜ 0.01) or the negative control group (96.40 ± 2.07; t =-24.19142,P ＜ 0.01).The experimental group showed slightly,but not significantly,lower cell proliferation (0.644 ± 0.027) than either the blank control group (0.669 ± 0.022; t=-1.60371,P＞ 0.05)or the negative control group (0.678 ± 0.027; t=-1.97828,P ＞ 0.05).In addition,there were no obvious differences between the three groups in the amounts of cells arrested in either the G1 phase or the S phase.Conclusion KiSS-1 overexpression suppresses the adhesion,invasion and motility,but not the proliferation,of hepatoma carcinoma cells in vitro.These findings imply that KiSS-1 might represent a promising new candidate for gene therapy against human hepatocellular carcinoma.
