中国医师杂志
中國醫師雜誌
중국의사잡지
JOURNAL OF CHINESE PHYSICIAN
2010年
8期
1009-1013
,共5页
RNA,小分子干扰%基因沉默%血管内皮生长因子C/遗传学%肠肿瘤/代谢/病理学%淋巴转移%淋巴管生成
RNA,小分子榦擾%基因沉默%血管內皮生長因子C/遺傳學%腸腫瘤/代謝/病理學%淋巴轉移%淋巴管生成
RNA,소분자간우%기인침묵%혈관내피생장인자C/유전학%장종류/대사/병이학%림파전이%림파관생성
RNA,small interfering%Gene silencing%Vascular endothelial growth factor C/GE%Intestinal neoplasms/ME/PA%Lymphatic metastasis%Lymphangiogenesis
目的 观察RNA干扰(RNA interference,RNAi)技术抑制大肠癌细胞株Lovo中VEGF-C基因的表达,探讨抑制shRNA-VEGF-C对人大肠癌细胞Lovo生物学特性的影响.方法 构建表达ShRNA-VEGF-C重组质粒转染Lovo细胞,72 h后用实时RT-PCR(real-time polymerase chain recation)检测VEGF-C mRNA表达;建立Lovo细胞皮下移植瘤裸鼠模型,注射shRNA-VEGF-C,动态观察肿瘤体积并于4周后处死裸鼠称取瘤重、计算局部淋巴结转移率,免疫组化法检测大肠癌组织微淋巴管密度(microlymphatic density,MLD).结果 转染shRNA-VEGF-C后,Lovo细胞VEGF-C mRNA表达下调;体内实验结果显示,4周后shRNA-VEGF-C组移植瘤体积[(324.9±64.8)mm3]明显小于空质粒组[(553.5±90.1)mm3]和生理盐水对照组[(570.1±85.4)mm3](P<0.01);shRNA-VEGF-C组瘤重[(3.01±0.55)g]也低于空质粒组[(4.65±0.65)g]和生理盐水对照组[(4.75±0.55)g](P<0.01);shRNA-VEGF-C组微淋巴管密度LMVD(15.5±6.90)明显低于HK组(24.18±6.45)和生理盐水对照组(29.59±8.21)(P<0.01);shRNA-VEGF-C组局部淋巴结转移率(30.1%)明显低于空质粒组(50.2%)和生理盐水组(53.1%).结论 shRNA-VEGF-C可影响VEGF-C诱导的淋巴管生成并抑制大肠癌淋巴结转移.
目的 觀察RNA榦擾(RNA interference,RNAi)技術抑製大腸癌細胞株Lovo中VEGF-C基因的錶達,探討抑製shRNA-VEGF-C對人大腸癌細胞Lovo生物學特性的影響.方法 構建錶達ShRNA-VEGF-C重組質粒轉染Lovo細胞,72 h後用實時RT-PCR(real-time polymerase chain recation)檢測VEGF-C mRNA錶達;建立Lovo細胞皮下移植瘤裸鼠模型,註射shRNA-VEGF-C,動態觀察腫瘤體積併于4週後處死裸鼠稱取瘤重、計算跼部淋巴結轉移率,免疫組化法檢測大腸癌組織微淋巴管密度(microlymphatic density,MLD).結果 轉染shRNA-VEGF-C後,Lovo細胞VEGF-C mRNA錶達下調;體內實驗結果顯示,4週後shRNA-VEGF-C組移植瘤體積[(324.9±64.8)mm3]明顯小于空質粒組[(553.5±90.1)mm3]和生理鹽水對照組[(570.1±85.4)mm3](P<0.01);shRNA-VEGF-C組瘤重[(3.01±0.55)g]也低于空質粒組[(4.65±0.65)g]和生理鹽水對照組[(4.75±0.55)g](P<0.01);shRNA-VEGF-C組微淋巴管密度LMVD(15.5±6.90)明顯低于HK組(24.18±6.45)和生理鹽水對照組(29.59±8.21)(P<0.01);shRNA-VEGF-C組跼部淋巴結轉移率(30.1%)明顯低于空質粒組(50.2%)和生理鹽水組(53.1%).結論 shRNA-VEGF-C可影響VEGF-C誘導的淋巴管生成併抑製大腸癌淋巴結轉移.
목적 관찰RNA간우(RNA interference,RNAi)기술억제대장암세포주Lovo중VEGF-C기인적표체,탐토억제shRNA-VEGF-C대인대장암세포Lovo생물학특성적영향.방법 구건표체ShRNA-VEGF-C중조질립전염Lovo세포,72 h후용실시RT-PCR(real-time polymerase chain recation)검측VEGF-C mRNA표체;건립Lovo세포피하이식류라서모형,주사shRNA-VEGF-C,동태관찰종류체적병우4주후처사라서칭취류중、계산국부림파결전이솔,면역조화법검측대장암조직미림파관밀도(microlymphatic density,MLD).결과 전염shRNA-VEGF-C후,Lovo세포VEGF-C mRNA표체하조;체내실험결과현시,4주후shRNA-VEGF-C조이식류체적[(324.9±64.8)mm3]명현소우공질립조[(553.5±90.1)mm3]화생리염수대조조[(570.1±85.4)mm3](P<0.01);shRNA-VEGF-C조류중[(3.01±0.55)g]야저우공질립조[(4.65±0.65)g]화생리염수대조조[(4.75±0.55)g](P<0.01);shRNA-VEGF-C조미림파관밀도LMVD(15.5±6.90)명현저우HK조(24.18±6.45)화생리염수대조조(29.59±8.21)(P<0.01);shRNA-VEGF-C조국부림파결전이솔(30.1%)명현저우공질립조(50.2%)화생리염수조(53.1%).결론 shRNA-VEGF-C가영향VEGF-C유도적림파관생성병억제대장암림파결전이.
Objective This study was to explore the inhibitory effect of shRNA-VEGF - C on growth of human colon cancer cell line Lovo in vitro and vivo. Methods Recombinant VEGF-C short hairpin RNA (shRNA) plasmid was constructed and transfected into Lovo cells. The expression of VEGF-C was detected at mRNA and protein levels by real-time reverse transcription-polymerase chain reaction (RT-PCR). In vivo study, xenograft tumors were established by injecting LOVO cells into nude mice, then shR-NA-VEGF-C were injected into the tumors, the tumor volume and weight and the incidences of lymph node metastasis were detected. Immunohistochemical staining was used to detect the lymphatic microvessel density of colon cancer tissues. Results After transfection of shRNA-VEGF-C, the mRNA of VEGF-C in Lovo cells were down-regulated. Four weeks after injection, the tumor volume and tumor weight in VEGF-C-shR-NA group were significantly smaller than that in empty plasmid group and NS group [(324. 9 ± 64. 8 ) mm3 vs. (553.5±90. 1)mm3 and (570. 1±85.4)mm3; (3.01 ±0.55)g vs (4.65 ±0.65)g and (4.75 ±0. 75)g]. The incidences of lymph node metastasis (30. 1% ) were significantly inhibited compared with empty plasmid group (50. 2% ) and normal saline group (53. 1% ). In shRNA-VEGF-C group, and microlymphatic density (15.5 ± 6. 90) was also decreased compared with empty plasmid group (24. 18 ±6. 45 ), and normal saline group (29. 59 ± 8. 21 ) ( all P <0. 01 ). Conclusion shRNA-VEGF-C can inhibit the growth of LOVO cells in vitro and vivo. VEGF-C may inhibit the lymph node metastasis of colon cancer by suppressing lymphangiogenesis.
