中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2015年
4期
711-714
,共4页
许践刚%林峰%郑双%沈贤%林刻智
許踐剛%林峰%鄭雙%瀋賢%林刻智
허천강%림봉%정쌍%침현%림각지
趋化因子%CXC趋化因子配体14%结肠癌%慢病毒%细胞增殖%细胞周期
趨化因子%CXC趨化因子配體14%結腸癌%慢病毒%細胞增殖%細胞週期
추화인자%CXC추화인자배체14%결장암%만병독%세포증식%세포주기
Chemokine%CXC ligand 14%Colon cancer%Lentivirus%Cell proliferation%Cell cycle
目的 观察CXC趋化因子配体14(CXCL14)在结肠癌组织中的表达,并探讨其对结肠癌细胞增殖及细胞周期的影响.方法 采用免疫组织化学法分析40例结肠癌及癌旁组织CXCL14蛋白的表达水平;构建含人CXCL14全长序列的pLenti6.3_CXCL14_ IRES2-EGFP重组质粒与包装质粒共转染293T细胞,制备表达CXCL14的慢病毒,转染HT29结肠癌细胞;通过Western blot检测转染后HT29结肠癌细胞中CXCL14的蛋白表达,采用细胞计数试剂盒(CCK-8)检测其对结肠癌细胞增殖能力的影响,最后采用流式细胞技术检测CXCL14干预后细胞周期分布的改变.结果 在结肠癌组织中,CXCL14蛋白表达较癌旁组织显著降低(P＜0.01),其平均吸光度值分别为0.541 1和0.1769,且其表达水平随着肿瘤分化程度降低而减少(P＜0.05).CXCL14慢病毒转染的HT29结肠癌细胞能明显上调CXCL14表达.CCK-8细胞增殖试验显示,CXCL14慢病毒转染的HT29结肠癌细胞活力明显受到抑制(P＜0.01).细胞周期分析显示,CXCL14干预组在G1期的细胞百分数[(67.46±0.92)％]显著低于对照组[(82.34±0.75)％,P＜0.05],S期的细胞百分数[(36.47±0.59)％]显著高于对照组[(21.97±0.64)％,P＜0.05].结论 结肠癌中的CXCL14可直接作为一个潜在的抑制基因参与结肠癌发生、发展过程.
目的 觀察CXC趨化因子配體14(CXCL14)在結腸癌組織中的錶達,併探討其對結腸癌細胞增殖及細胞週期的影響.方法 採用免疫組織化學法分析40例結腸癌及癌徬組織CXCL14蛋白的錶達水平;構建含人CXCL14全長序列的pLenti6.3_CXCL14_ IRES2-EGFP重組質粒與包裝質粒共轉染293T細胞,製備錶達CXCL14的慢病毒,轉染HT29結腸癌細胞;通過Western blot檢測轉染後HT29結腸癌細胞中CXCL14的蛋白錶達,採用細胞計數試劑盒(CCK-8)檢測其對結腸癌細胞增殖能力的影響,最後採用流式細胞技術檢測CXCL14榦預後細胞週期分佈的改變.結果 在結腸癌組織中,CXCL14蛋白錶達較癌徬組織顯著降低(P＜0.01),其平均吸光度值分彆為0.541 1和0.1769,且其錶達水平隨著腫瘤分化程度降低而減少(P＜0.05).CXCL14慢病毒轉染的HT29結腸癌細胞能明顯上調CXCL14錶達.CCK-8細胞增殖試驗顯示,CXCL14慢病毒轉染的HT29結腸癌細胞活力明顯受到抑製(P＜0.01).細胞週期分析顯示,CXCL14榦預組在G1期的細胞百分數[(67.46±0.92)％]顯著低于對照組[(82.34±0.75)％,P＜0.05],S期的細胞百分數[(36.47±0.59)％]顯著高于對照組[(21.97±0.64)％,P＜0.05].結論 結腸癌中的CXCL14可直接作為一箇潛在的抑製基因參與結腸癌髮生、髮展過程.
목적 관찰CXC추화인자배체14(CXCL14)재결장암조직중적표체,병탐토기대결장암세포증식급세포주기적영향.방법 채용면역조직화학법분석40례결장암급암방조직CXCL14단백적표체수평;구건함인CXCL14전장서렬적pLenti6.3_CXCL14_ IRES2-EGFP중조질립여포장질립공전염293T세포,제비표체CXCL14적만병독,전염HT29결장암세포;통과Western blot검측전염후HT29결장암세포중CXCL14적단백표체,채용세포계수시제합(CCK-8)검측기대결장암세포증식능력적영향,최후채용류식세포기술검측CXCL14간예후세포주기분포적개변.결과 재결장암조직중,CXCL14단백표체교암방조직현저강저(P＜0.01),기평균흡광도치분별위0.541 1화0.1769,차기표체수평수착종류분화정도강저이감소(P＜0.05).CXCL14만병독전염적HT29결장암세포능명현상조CXCL14표체.CCK-8세포증식시험현시,CXCL14만병독전염적HT29결장암세포활력명현수도억제(P＜0.01).세포주기분석현시,CXCL14간예조재G1기적세포백분수[(67.46±0.92)％]현저저우대조조[(82.34±0.75)％,P＜0.05],S기적세포백분수[(36.47±0.59)％]현저고우대조조[(21.97±0.64)％,P＜0.05].결론 결장암중적CXCL14가직접작위일개잠재적억제기인삼여결장암발생、발전과정.
Objective To investigate the expression of CXC ligand 14 (CXCL14) in colon cancer tissues and evaluate the direct effect of CXCL14 overexpression on cell proliferation and cycle.Methods The recombinant plasmid of pLenti6.3_CXCL14_IRES2-EGFP encoding the CXCL14 gene and packaging plasmid mix then was transformed into 293T cells to obtain replication defective lentivirus expressing CXCL14 and enhanced green fluorescence protein (EGFP).After colon cancer HT29 cells were treated with lentivirus,overexpression of CXCL14 was then detected by Western blotting.Cell viability was assayed using cell counting kit-8 (CCK-8),and cell cycle distributions were detected by flow cytometry analysis.Results Our data indicated the level of CXCL14 protein was declined in colon cancer tissues compared to the paired normal tissues (P＜0.01).The mean absorbance values of CXCL14 in tumor and normal specimen were 0.541 1 and 0.176 9respectively.Meanwhile,the expression of CXCL14 was declined as the decrease of tumor differentiation degree (P ＜ 0.05).The transfection of lentivirus encoding the CXCL14 gene could significantly upregulate the expression of CXCL14 protein in colon cells.The result of CCK-8 demonstrated that overexpression of CXCL14 in transfected colon cells had an inhibitory effect on cell proliferation (P ＜0.01).Additionally,cell cycle alternations by the analysis of flow cytometry indicated a significant decrease in G1 phase arrest in CXCL14-letivirus-infected HT29 cells [(67.46 ± 0.92) ％] as compared with contrl group [(82.34 ± 0.75) ％,P ＜ 0.05],accompanied by a significant increasein Sphase [(36.47±0.59)％ vs.(21.97±0.64)％,P＜0.05].Condusion CXCL14 may play a pivotal role as a potential tumor suppressor in the development of colon cancer.
