中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
Chinese Journal of Experimental Surgery
2015年
10期
2347-2349
,共3页
李云%郑建伟%张晓梅%易继林%秦仁义%吴在德%薛新波%申铭
李雲%鄭建偉%張曉梅%易繼林%秦仁義%吳在德%薛新波%申銘
리운%정건위%장효매%역계림%진인의%오재덕%설신파%신명
肿瘤抑制因子-1%肝细胞癌%黏附%侵袭转移
腫瘤抑製因子-1%肝細胞癌%黏附%侵襲轉移
종류억제인자-1%간세포암%점부%침습전이
Tumor suppressor in lung cancer-1%Human hepatocellular carcinoma%Adhesion%Invasion and metastasis
目的 观察肺癌肿瘤抑制因子-1(TSLC-1)对人肝癌细胞黏附及运动功能作用的影响,探讨TSLC-1基因在转移中的作用机制.方法 提取正常肝细胞株L02、肝癌细胞低侵袭细胞株MHCC97-L、肝癌细胞高侵袭细胞株MHCC97-H总RNA,反转录合成第1链cDNA,对反转录产物进行荧光定量聚合酶链反应(FQ-PCR).用管家基因β-肌动蛋白(β-actin)的cDNA起始浓度作为参照标准化TSLC-1的cDNA起始浓度,对结果进行单因素方差分析.用重组真核表达载体pcDNA3.1 hisC-TSLC-1转染7402细胞并经过稳定筛选,采用细胞计数法检测TSLC-1基因对7402细胞同质黏附、异质黏附作用的影响,细胞划痕实验检测TSLC-1基因对7402细胞运动能力的影响.结果 正常肝细胞L02、肝癌细胞MHCC97-L、MHCC97-H标准化后的TSLC-1 cDNA起始浓度分别为(33.15±0.05)×103、(9.06±0.03)×103、(6.14±0.09)×103,各细胞株之间TSLC-1表达量差异有统计学意义(P＜0.01).TSLC-1基因转染7402细胞后,同质黏附作用后细胞数[(1301 ±24)个]高于空质粒转染组[(1 279±20)个]和对照组[(987 ±26)个],异质黏附作用后实验组细胞运动迁移能力明显低于空质粒转染组和对照组,组间差异有统计学意义(P＜0.05).结论 TSLC-1基因表达可使人正常肝细胞7402同质黏附作用降低,异质黏附作用降低,运动迁移能力降低.因此,TSLC-1基因具有抑制肝癌细胞发生转移的功能,其机制是改变了细胞基质及细胞间的黏附能力、运动能力,增加TSLC-1基因的表达可能成为一个新的潜在的治疗肝癌的分子靶点.
目的 觀察肺癌腫瘤抑製因子-1(TSLC-1)對人肝癌細胞黏附及運動功能作用的影響,探討TSLC-1基因在轉移中的作用機製.方法 提取正常肝細胞株L02、肝癌細胞低侵襲細胞株MHCC97-L、肝癌細胞高侵襲細胞株MHCC97-H總RNA,反轉錄閤成第1鏈cDNA,對反轉錄產物進行熒光定量聚閤酶鏈反應(FQ-PCR).用管傢基因β-肌動蛋白(β-actin)的cDNA起始濃度作為參照標準化TSLC-1的cDNA起始濃度,對結果進行單因素方差分析.用重組真覈錶達載體pcDNA3.1 hisC-TSLC-1轉染7402細胞併經過穩定篩選,採用細胞計數法檢測TSLC-1基因對7402細胞同質黏附、異質黏附作用的影響,細胞劃痕實驗檢測TSLC-1基因對7402細胞運動能力的影響.結果 正常肝細胞L02、肝癌細胞MHCC97-L、MHCC97-H標準化後的TSLC-1 cDNA起始濃度分彆為(33.15±0.05)×103、(9.06±0.03)×103、(6.14±0.09)×103,各細胞株之間TSLC-1錶達量差異有統計學意義(P＜0.01).TSLC-1基因轉染7402細胞後,同質黏附作用後細胞數[(1301 ±24)箇]高于空質粒轉染組[(1 279±20)箇]和對照組[(987 ±26)箇],異質黏附作用後實驗組細胞運動遷移能力明顯低于空質粒轉染組和對照組,組間差異有統計學意義(P＜0.05).結論 TSLC-1基因錶達可使人正常肝細胞7402同質黏附作用降低,異質黏附作用降低,運動遷移能力降低.因此,TSLC-1基因具有抑製肝癌細胞髮生轉移的功能,其機製是改變瞭細胞基質及細胞間的黏附能力、運動能力,增加TSLC-1基因的錶達可能成為一箇新的潛在的治療肝癌的分子靶點.
목적 관찰폐암종류억제인자-1(TSLC-1)대인간암세포점부급운동공능작용적영향,탐토TSLC-1기인재전이중적작용궤제.방법 제취정상간세포주L02、간암세포저침습세포주MHCC97-L、간암세포고침습세포주MHCC97-H총RNA,반전록합성제1련cDNA,대반전록산물진행형광정량취합매련반응(FQ-PCR).용관가기인β-기동단백(β-actin)적cDNA기시농도작위삼조표준화TSLC-1적cDNA기시농도,대결과진행단인소방차분석.용중조진핵표체재체pcDNA3.1 hisC-TSLC-1전염7402세포병경과은정사선,채용세포계수법검측TSLC-1기인대7402세포동질점부、이질점부작용적영향,세포화흔실험검측TSLC-1기인대7402세포운동능력적영향.결과 정상간세포L02、간암세포MHCC97-L、MHCC97-H표준화후적TSLC-1 cDNA기시농도분별위(33.15±0.05)×103、(9.06±0.03)×103、(6.14±0.09)×103,각세포주지간TSLC-1표체량차이유통계학의의(P＜0.01).TSLC-1기인전염7402세포후,동질점부작용후세포수[(1301 ±24)개]고우공질립전염조[(1 279±20)개]화대조조[(987 ±26)개],이질점부작용후실험조세포운동천이능력명현저우공질립전염조화대조조,조간차이유통계학의의(P＜0.05).결론 TSLC-1기인표체가사인정상간세포7402동질점부작용강저,이질점부작용강저,운동천이능력강저.인차,TSLC-1기인구유억제간암세포발생전이적공능,기궤제시개변료세포기질급세포간적점부능력、운동능력,증가TSLC-1기인적표체가능성위일개신적잠재적치료간암적분자파점.
Objective To study the effecof tumosuppressoin lung cancer-1 (TSLC-1) gene transfected into hepatocellulacarcinomcell line 7402, and to explore the mechanism of TSLC-1 gene on the metastaseof human hepatocellulacarcinom(HCC) cells.MethodThe recombinanplasmid of pcDNA3.1 hisC-TSLC-1 wastably transfected into 7402 cell.The effecto 7402 gene on the homogeneity adhesion and the heterogeneity adhesion of hepatocellulacarcinomcellwere measured with cell count, The in vitro migration of pcDNA3.1hisC-TSLC-1 and mock cellon extrcellulamatrix natrigel waassayed by monolayewound healing test.ResultThe normalized initial cDNconcentration of TSLC-1 in L02, MHCC97-L and MHCC97-H were (33.15 ± 0.05) × 103, (9.06 ± 0.03) × 103,(6.14 ±0.09) × 103, respectively.There wasignificandifference in expression level in livecancecell strainwith differenmetastatipotential.The homogeneity adhesion of 7402-TSLC-1 cell group (1 301 ± 24) wahighethan empty vectogroup (1 279 ± 20) and contrasgroup (987 ± 26), and the heterogeneity adhesion walowethan empty vectogroup and contrasgroup.The in vitro cell adhesion experimenshowed thathe exogenouTSLC-1 gene in the 7402-TSLC-1 celldecreased cell adhesion to matrigel and fibnectin extrcellulamatrix by 49.3％ and 37.5％, respectively, compared with the control cells.The migration of the 7402-TSLC-1 cellon the matrigel waraised by 31.5％.Conclusion The human imprinted gene TSLC-1 can reduce homogeneity adhesion, promote heterogeneity adhesion and migration of 7402-TSLC-1 cells.TSLC-1 gene mighfunction ametastasisuppressogene foHCC.
