基础医学与临床
基礎醫學與臨床
기출의학여림상
Basic & Clinical Medicine
2015年
1期
54-59
,共6页
邓大炜%孔宪炳%王平%于庆三%王强
鄧大煒%孔憲炳%王平%于慶三%王彊
산대위%공헌병%왕평%우경삼%왕강
Gli2%上皮间质转化( EMT)%SMMC-7721%侵袭%黏附
Gli2%上皮間質轉化( EMT)%SMMC-7721%侵襲%黏附
Gli2%상피간질전화( EMT)%SMMC-7721%침습%점부
Gli2%epithelial-mesenchymal transition (EMT)%SMMC-7721 cell%invasion%adhesion
目的：探讨胶质瘤相关癌基因同源蛋白2（Gli2）对肝癌细胞系SMMC-7721上皮间质转化（EMT）和侵袭能力的影响及机制。方法合成shRNA-Gli2、shRNA-NC病毒载体，实验设shRNA-Gli2组、shRNA-NC组和空白组，转染SMMC-7721细胞。用Transwell小室和细胞黏附实验观察细胞侵袭及黏附能力；用qRT-PCR和Western blot检测细胞间Gli2、E-cadherin、N-cadherin及vimentin基因和蛋白的表达。结果在不同肝癌细胞系中，随着肝癌细胞系侵袭能力的增强，Gli2的表达逐渐增高，shRNA-Gli2组与对照组相比侵袭能力降低，同种黏附能力增加，异种黏附能力降低（P＜0.05）。 shRNA-Gli2组与对照组相比，E-cadherin 表达明显增高而N-cadherin 和vimentin 表达明显降低（ P＜0.05）。结论沉默Gli2的表达能够促进肝癌细胞SMMC-7721 EMT的反转，并降低其侵袭能力，机制可能与E-cadherin的上调和N-cadherin、vimentin的下调有关。
目的：探討膠質瘤相關癌基因同源蛋白2（Gli2）對肝癌細胞繫SMMC-7721上皮間質轉化（EMT）和侵襲能力的影響及機製。方法閤成shRNA-Gli2、shRNA-NC病毒載體，實驗設shRNA-Gli2組、shRNA-NC組和空白組，轉染SMMC-7721細胞。用Transwell小室和細胞黏附實驗觀察細胞侵襲及黏附能力；用qRT-PCR和Western blot檢測細胞間Gli2、E-cadherin、N-cadherin及vimentin基因和蛋白的錶達。結果在不同肝癌細胞繫中，隨著肝癌細胞繫侵襲能力的增彊，Gli2的錶達逐漸增高，shRNA-Gli2組與對照組相比侵襲能力降低，同種黏附能力增加，異種黏附能力降低（P＜0.05）。 shRNA-Gli2組與對照組相比，E-cadherin 錶達明顯增高而N-cadherin 和vimentin 錶達明顯降低（ P＜0.05）。結論沉默Gli2的錶達能夠促進肝癌細胞SMMC-7721 EMT的反轉，併降低其侵襲能力，機製可能與E-cadherin的上調和N-cadherin、vimentin的下調有關。
목적：탐토효질류상관암기인동원단백2（Gli2）대간암세포계SMMC-7721상피간질전화（EMT）화침습능력적영향급궤제。방법합성shRNA-Gli2、shRNA-NC병독재체，실험설shRNA-Gli2조、shRNA-NC조화공백조，전염SMMC-7721세포。용Transwell소실화세포점부실험관찰세포침습급점부능력；용qRT-PCR화Western blot검측세포간Gli2、E-cadherin、N-cadherin급vimentin기인화단백적표체。결과재불동간암세포계중，수착간암세포계침습능력적증강，Gli2적표체축점증고，shRNA-Gli2조여대조조상비침습능력강저，동충점부능력증가，이충점부능력강저（P＜0.05）。 shRNA-Gli2조여대조조상비，E-cadherin 표체명현증고이N-cadherin 화vimentin 표체명현강저（ P＜0.05）。결론침묵Gli2적표체능구촉진간암세포SMMC-7721 EMT적반전，병강저기침습능력，궤제가능여E-cadherin적상조화N-cadherin、vimentin적하조유관。
Objective To explore the effect and mechanism of Gli2 on EMT and invasion in the hepatocellular car-cinoma cell line SMMC-7721.Methods shRNA of Gli2 and Negative control (NC) shRNA were constructed and transfected into SMMC-7721 cells.shRNA-Gli2 group,shRNA-NC group and blank group were set up .Transwell chambers assay , adhesion experiments were used to detect the ability of invasion ,homogeneous and heterogeneous cells intercellular adhesion of each group .Meanwhile, the qRT-PCR, Western blot were used to examine Gli2, E-cadherin ,N-cadherin and vimentin mRNA and protein expression .Results In different hepatocellular carcinoma cell lines and be along with the increasing ability of the invasion in hepatocellular carcinoma cell lines , Gli2 expres-sion was higher .Compared with the shRNA-NC group and blank control group ,the interfered group extensive cells invasion ability was inhibited ( P <0.05 ) , homogeneous cells intercellular adhesion increased and heterogeneous cells intercellular was opposite ( P <0.05 ) , meanwhile , the expression of E-cadherin was declined significantly ( P<0.05 ) ,the expression of N-cadherin , vimentin raised significantly ( P<0.05 ) .Conclusions Silencing Gli2 gene can reverse the EMT of the hepatocellular carcinoma cell line SMMC-7721 and inhibit cell invasion ,its mecha- <br> nism may be related to the up-regulation of E-cadherin and the dow N-regulation of N-cadherin, vimentin.
