CAJ | 학술논문

目的探讨细胞密度对结肠癌细胞整合素αγβ6表达和基质会属蛋白酶9(MMP-9)分泌的影响. 方法用流式细胞仪榆测结肠癌细胞系WiDr和人正常角质细胞系HaCaT细胞株在高、低细胞密度培养条件下αγβ6表达的情况;用Biotrak MMP-9测定仪和明胶酶谱法分别测定不同的结肠癌细胞WiDr和SW480细胞株在高、低密度培养条件下MMP-9的活性和分泌水平. 结果表达αγβ6的结肠癌WiDr细胞出现细胞密度依赖的αγβ6表达增加,而正常角质细胞系HaCaT细胞在高、低密度培养条件下αγβ6表达无改变.Biotrak MMP-9活性分析显示,每个表达αγβ6的WiDr细胞和SW480β6细胞的MMP-9分泌量在高密度培养条件下分别是(3.3±1.2)×10-7-ng和(27.2±3.0)×10-7ng,在低密度培养条件下分别足(1.8±0.7)×10-7ng和(10.9±2.0)×10-7ng,而每个缺乏αγβ6表达的SW480 wild细胞在高、低细胞密度培养条件下分别足(3.9±1.7)×10-7ng和(3.8±0.7)×10-7ng,MMP-9分泌水平无明显变化(t=0.47,P＞0.05).明胶酶谱分析显示SW480 β6细胞的MMP-9活性水平在高密度培养时比低密度培养明显增加,而SW480 wild和HaCaT细胞的MMP-9活性水平无明显改变.结论细胞高密度诱导结肠癌细胞αγβ6表达,促进MMP-9的分泌,构成了癌细胞永生化侵袭浸润性生长的基础.
목적 탐토세포밀도대결장암세포정합소αγβ6표체화기질회속단백매9(MMP-9)분비적영향. 방법용류식세포의유측결장암세포계WiDr화인정상각질세포계HaCaT세포주재고、저세포밀도배양조건하αγβ6표체적정황;용Biotrak MMP-9측정의화명효매보법분별측정불동적결장암세포WiDr화SW480세포주재고、저밀도배양조건하MMP-9적활성화분비수평. 결과표체αγβ6적결장암WiDr세포출현세포밀도의뢰적αγβ6표체증가,이정상각질세포계HaCaT세포재고、저밀도배양조건하αγβ6표체무개변.Biotrak MMP-9활성분석현시,매개표체αγβ6적WiDr세포화SW480β6세포적MMP-9분비량재고밀도배양조건하분별시(3.3±1.2)×10-7-ng화(27.2±3.0)×10-7ng,재저밀도배양조건하분별족(1.8±0.7)×10-7ng화(10.9±2.0)×10-7ng,이매개결핍αγβ6표체적SW480 wild세포재고、저세포밀도배양조건하분별족(3.9±1.7)×10-7ng화(3.8±0.7)×10-7ng,MMP-9분비수평무명현변화(t=0.47,P＞0.05).명효매보분석현시SW480 β6세포적MMP-9활성수평재고밀도배양시비저밀도배양명현증가,이SW480 wild화HaCaT세포적MMP-9활성수평무명현개변.결론 세포고밀도유도결장암세포αγβ6표체,촉진MMP-9적분비,구성료암세포영생화침습침윤성생장적기출.
Objective To detect the effect of cell density off both integrin αγβ6 expression and matrix metalloproteinase-9(MMP-9)secretion in colon cancer cells. Methods Flow cytometry was applied to analyze αγβ6 expression in human WiDr colon cancer cell lines and human HaCaT keratinocyte cells, respectively,at high-and low-cell density culture.The MMP-9 aetivity level for various coloIl cancer cell lines, WiDr and SW480 cells at high-and low-cell density culture was analyzed using Biotrak MMP-9 activity assay and Gelatin Zymography assay, respectively. Results High cell density significantly enhances integrin αγβ6 expression for WiDr cells expressing αγβ6 compared with low density,but no increase was observed for human keratinocyte HaCaT cells. Biotrak MMP-9 assay indicated that the amount of MMP-9 secreted per cell for WiDr and SW480 B6 cells at high cell density culture was(3.3±1.2)×10-7ng/cell and(27.2±3.0)× 10-7ng/cell respectively; However,at low cell density it was(1.8±0.7)× 10-7ng/cell and(10.9±2.0)×10-7 ng/cell,respectively. It was 2-3-fold higher for WiDr and SW480 β6 cells at high cell density compared with that at low cell density, but no density-dependent increase observed for SW480 wild cells lack αγβ6 expression(t=0.47,P＞0.05),MMP-9 secretion for SW480 wild cells was(3.9±1.7)× 10-7 ng/cell at hish cell density and(3.8 ±0.7)×10-7 ng/cell at low eell density(P＞0.05),respectively. Gelatin zymography assay also indicated that the level of MMP-9 in SW480 B6 cells expressing αγβ6 was evidently higher at high density than at low density, however no density-dependent increase observed for SW480 wild cells and HaCaT cells. Conclusions High cell density induces integrin αγβ6 expression and promotes MMP-9 secretion in colon cancer cells, which constitutes the basis for a self-perpetuating system of tumor infiltrating growth in colon cancer progression.