中国医师杂志
中國醫師雜誌
중국의사잡지
JOURNAL OF CHINESE PHYSICIAN
2015年
4期
528-532
,共5页
朱海宏%吴新民%叶谢智华
硃海宏%吳新民%葉謝智華
주해굉%오신민%협사지화
胃肿瘤/代谢%基质金属蛋白酶11/代谢%Sp转录因子类/代谢
胃腫瘤/代謝%基質金屬蛋白酶11/代謝%Sp轉錄因子類/代謝
위종류/대사%기질금속단백매11/대사%Sp전록인자류/대사
Stomach neoplasms/ME%Matrix metalloproteinase 11/ME%Sp transcription factors/ME
目的 探讨基质金属蛋白酶11(MMP-11)在胃癌细胞的表达及其与转录因子Sp1的相关性.方法 以10个胃癌细胞系为对象,采用RT-PCR、differential PCR、Western-Blot并结合DNA测序,检测了MMP-11及SP1的基因表达情况.结果 MMP-11在胃癌细胞系差异表达,通过RT-PCR及差异性PCR,观察到34循环下SNU16、BGC823、MKN45、SGC7901、MGC803、SNU5、PAMC82中MMP-11表达灰度比分别是:3.32 ±0.12、2.22±0.11、1.41±0.12、1.35±0.05、1.19±0.05、0.97±0.05、0.79±0.05(F=371.54,P＜0.001).SNU16细胞MMP-11 DNA片段表达灰度比是其它细胞系的2～2.5倍,SNU16在MMP-11 DNA的拷贝数上较其它细胞系呈倍数关系.说明SNU16细胞系存在染色体扩增.不同胃癌细胞系细胞总蛋白提取物中Sp1蛋白、MMP-11蛋白有显著表达,二者之间具有相关表达趋势(r =0.951,P＜0.05).结论 Sp1蛋白、MMP-11蛋白在胃癌细胞系中显著表达,正常组织低表达或不表达,而且二者具有相关性.为确定Sp1通过MMP-11基因5'端上游区GC-box对MMP-11进行调节的研究奠定了基础.
目的 探討基質金屬蛋白酶11(MMP-11)在胃癌細胞的錶達及其與轉錄因子Sp1的相關性.方法 以10箇胃癌細胞繫為對象,採用RT-PCR、differential PCR、Western-Blot併結閤DNA測序,檢測瞭MMP-11及SP1的基因錶達情況.結果 MMP-11在胃癌細胞繫差異錶達,通過RT-PCR及差異性PCR,觀察到34循環下SNU16、BGC823、MKN45、SGC7901、MGC803、SNU5、PAMC82中MMP-11錶達灰度比分彆是:3.32 ±0.12、2.22±0.11、1.41±0.12、1.35±0.05、1.19±0.05、0.97±0.05、0.79±0.05(F=371.54,P＜0.001).SNU16細胞MMP-11 DNA片段錶達灰度比是其它細胞繫的2～2.5倍,SNU16在MMP-11 DNA的拷貝數上較其它細胞繫呈倍數關繫.說明SNU16細胞繫存在染色體擴增.不同胃癌細胞繫細胞總蛋白提取物中Sp1蛋白、MMP-11蛋白有顯著錶達,二者之間具有相關錶達趨勢(r =0.951,P＜0.05).結論 Sp1蛋白、MMP-11蛋白在胃癌細胞繫中顯著錶達,正常組織低錶達或不錶達,而且二者具有相關性.為確定Sp1通過MMP-11基因5'耑上遊區GC-box對MMP-11進行調節的研究奠定瞭基礎.
목적 탐토기질금속단백매11(MMP-11)재위암세포적표체급기여전록인자Sp1적상관성.방법 이10개위암세포계위대상,채용RT-PCR、differential PCR、Western-Blot병결합DNA측서,검측료MMP-11급SP1적기인표체정황.결과 MMP-11재위암세포계차이표체,통과RT-PCR급차이성PCR,관찰도34순배하SNU16、BGC823、MKN45、SGC7901、MGC803、SNU5、PAMC82중MMP-11표체회도비분별시:3.32 ±0.12、2.22±0.11、1.41±0.12、1.35±0.05、1.19±0.05、0.97±0.05、0.79±0.05(F=371.54,P＜0.001).SNU16세포MMP-11 DNA편단표체회도비시기타세포계적2～2.5배,SNU16재MMP-11 DNA적고패수상교기타세포계정배수관계.설명SNU16세포계존재염색체확증.불동위암세포계세포총단백제취물중Sp1단백、MMP-11단백유현저표체,이자지간구유상관표체추세(r =0.951,P＜0.05).결론 Sp1단백、MMP-11단백재위암세포계중현저표체,정상조직저표체혹불표체,이차이자구유상관성.위학정Sp1통과MMP-11기인5'단상유구GC-box대MMP-11진행조절적연구전정료기출.
Objective To explore the expression of matrix metalloproteinase-11 (MMP-11) gene in gastric cancer cells and its correlation with transcription factor Sp1.Methods T In the present study,the expression of the MMP-11 was studied and the importance of Sp1 in the expression of the MMP-11 was proved with reverse transcription polymerase chain reaction (RT-PCR),differential PCR,Western-Blot,and DNA sequencing analysis in 10 kinds of the gastric cancer cell lines.Results MMP-11 cDNA was amplified with RT-PCR and differential PCR.In 34 cycle,the MMP-11/β-actin of SNU16,BGC823,MKN45,SGC7901,MGC803,SNU5 and PAMC82 were 3.32 ±0.12,2.22 ±0.11,1.41 ±0.12,1.35 ± 0.05,1.19 ± 0.05,0.97 ± 0.05,and 0.79 ± 0.05 (F =371.54,P ＜ 0.001).DNA sequencing analysis and BLAST showed the MMP11/β-actin of SNU16 DNA was 2 ～2.5 times as much as that of others.This result indicated that MMP-11 gene was amplified on the SNU16 DNA.Differential PCR and DNA sequencing analysis were carried out to amplify the MMP-11 gene in the SNU16 cell.Toward a better understanding of transcription of the MMP-1 1 gene,bioinformics technology was used to demonstrate the existence of two GC-boxes of the 5'-flanking region of MMP-11 DNA.Sp1 protein and MMP-11 protein were over-expressed in tumor cells and tended to have correlation with them (r =0.951,P ＜ 0.05).Conclusions The present results described the expressions of MMP-11 and Sp1 in gastric cancer cells.Moreover and clarified the correlation between MMP-11 and Sp1.It will establish the basic theory in order to confirm further whether Sp1 is a significant promoter of the 5'-flanking region of MMP-11 DNA.
