中国现代医生
中國現代醫生
중국현대의생
CHINA MODERN DOCTOR
2015年
15期
14-16,161
,共4页
王欢%袁武锋%蒋雷%费鲜明
王歡%袁武鋒%蔣雷%費鮮明
왕환%원무봉%장뢰%비선명
趋化因子受体CCR7%pLVX-Puro质粒%SGC7901细胞%胃癌%转移
趨化因子受體CCR7%pLVX-Puro質粒%SGC7901細胞%胃癌%轉移
추화인자수체CCR7%pLVX-Puro질립%SGC7901세포%위암%전이
Chemokine receptor CCR7%pLVX-Puro plasmid%SGC7901 cell%Stomach cancer%Metastasis
目的:构建pLVX-Puro-CCR7稳定表达系统并研究CCR7基因在胃癌转移中的作用。方法采用PCR方法扩增CCR7基因并构建pLVX-Puro-CCR7重组质粒,通过脂质体转染法瞬时转染SGC7901细胞株,嘌呤霉素对该细胞株进行筛选并建立稳定表达细胞株。 Western blot方法检测稳定转染的SGC7901细胞中CCR7表达情况。Transwell方法分析过度表达CCR7基因对SGC7901胃癌细胞体外迁移的影响。活体成像观察过度表达CCR7的SGC7901胃癌细胞株在裸鼠体内转移能力。结果成功构建pLVX-Puro-CCR7重组质粒并稳定转染SGC7901细胞株,该细胞株能够正确翻译及表达CCR7蛋白。过度表达CCR7基因的SGC7901胃癌细胞株体外迁移能力和在裸鼠体内转移能力显著增强(P<0.05)。结论 CCR7可能参与促进胃癌细胞转移的功能。
目的:構建pLVX-Puro-CCR7穩定錶達繫統併研究CCR7基因在胃癌轉移中的作用。方法採用PCR方法擴增CCR7基因併構建pLVX-Puro-CCR7重組質粒,通過脂質體轉染法瞬時轉染SGC7901細胞株,嘌呤黴素對該細胞株進行篩選併建立穩定錶達細胞株。 Western blot方法檢測穩定轉染的SGC7901細胞中CCR7錶達情況。Transwell方法分析過度錶達CCR7基因對SGC7901胃癌細胞體外遷移的影響。活體成像觀察過度錶達CCR7的SGC7901胃癌細胞株在裸鼠體內轉移能力。結果成功構建pLVX-Puro-CCR7重組質粒併穩定轉染SGC7901細胞株,該細胞株能夠正確翻譯及錶達CCR7蛋白。過度錶達CCR7基因的SGC7901胃癌細胞株體外遷移能力和在裸鼠體內轉移能力顯著增彊(P<0.05)。結論 CCR7可能參與促進胃癌細胞轉移的功能。
목적:구건pLVX-Puro-CCR7은정표체계통병연구CCR7기인재위암전이중적작용。방법채용PCR방법확증CCR7기인병구건pLVX-Puro-CCR7중조질립,통과지질체전염법순시전염SGC7901세포주,표령매소대해세포주진행사선병건립은정표체세포주。 Western blot방법검측은정전염적SGC7901세포중CCR7표체정황。Transwell방법분석과도표체CCR7기인대SGC7901위암세포체외천이적영향。활체성상관찰과도표체CCR7적SGC7901위암세포주재라서체내전이능력。결과성공구건pLVX-Puro-CCR7중조질립병은정전염SGC7901세포주,해세포주능구정학번역급표체CCR7단백。과도표체CCR7기인적SGC7901위암세포주체외천이능력화재라서체내전이능력현저증강(P<0.05)。결론 CCR7가능삼여촉진위암세포전이적공능。
Objective To construct stable expression system of pLVX-Pro-CCR7 and study the function of CCR7 gene in metastasis of stomach cancer. Methods PCR method was used to amplify CCR7 gene and construct pLVX-Puro-CCR7 recombinant plasmid. Lipofection was used to infect SGC7901 cell strain immediately, and puromycin was used to screen the cell strain and establish a cell strain with stable expression. Western blot method was used to test the ex-pression of CCR7 in SGC7901 cells which were stably infected. Transwell method was used to analyze the effect of overly expressed CCR7 gene on in vitro migration of stomach cancer cells of SGC7901. Living imaging was used to ob-serve the metastasis ability of stomach cancer cell strain of SGC7901 which overly expressed CCR7 in nude mice. Re-sults pLVX-Puro-CCR7 recombinant plasmid was successfully constructed and SGC791 cell strain was stably infected. The cell strain was able to correctly translate and express CCR7 protein. Metastasis ability in nude mice and in vitro migration ability of SGC7901 stomach cancer cell strain which overly expressed CCR7 gene significantly improved (P<0.05). Conclusion CCR7 may be involved in the function of promoting metastasis of stomach cancer cells.