CAJ | 학술논문

目的观察淋巴内皮细胞在血管内皮生长因子(VEGF)-C促进肠癌细胞生长过程中是否发挥作用.方法运用过表达VEGF-C(h-VEGF-C)和抑制VEGF-C(L-VEGF-C)表达的慢病毒分别感染CT26细胞株,Western blot验证慢病毒载体的有效性.然后将经过慢病毒处理过两种细胞株和未经处理的CT26细胞株进行裸鼠注射,注射部位为小鼠尾部直肠处,进行小鼠模型构建.动物模型构建成功后进行组织切片免疫荧光染色;染色指标为LYVE-1(标记微淋巴管)和CD31(标记血管密度).最后亚甲蓝染色观察小鼠肠道肿瘤数目及大小.结果 Western blot结果验证了过表达和抑制VEGF-C表达慢病毒有效性,免疫荧光结果显示过表达VEGF-C的裸鼠淋巴管密度明显高于空白对照组(9.10 ±0.50比5.82±0.81),差异有统计学意义(P＜0.01);抑制VEGF-C的裸鼠淋巴管密度明显低于空白对照组(2.62±0.41比5.80±0.81),差异有统计学意义(P＜0.01).同样地,过表达VEGF-C的裸鼠血管密度明显高于空白对照组(2.68±0.23比2.01±0.15),差异有统计学意义(P＜0.05);抑制VEGF-C的裸鼠血管密度明显低于空白对照组(2.01±0.15比1.05±0.47),差异有统计学意义(P＜0.05).同时,亚甲蓝染色结果显示过表达VEGF-C时小鼠肠道肿瘤数目明显多于抑制VEGF-C表达的小鼠(13.4±5.2比4.5±1.8;P＜0.01);而且两组之间肠道肿瘤大小也有明显差异.结论 VEGF-C可以通过调节淋巴内皮细胞进行促进肠癌细胞的生长.
목적 관찰림파내피세포재혈관내피생장인자(VEGF)-C촉진장암세포생장과정중시부발휘작용.방법 운용과표체VEGF-C(h-VEGF-C)화억제VEGF-C(L-VEGF-C)표체적만병독분별감염CT26세포주,Western blot험증만병독재체적유효성.연후장경과만병독처리과량충세포주화미경처리적CT26세포주진행라서주사,주사부위위소서미부직장처,진행소서모형구건.동물모형구건성공후진행조직절편면역형광염색;염색지표위LYVE-1(표기미림파관)화CD31(표기혈관밀도).최후아갑람염색관찰소서장도종류수목급대소.결과 Western blot결과험증료과표체화억제VEGF-C표체만병독유효성,면역형광결과현시과표체VEGF-C적라서림파관밀도명현고우공백대조조(9.10 ±0.50비5.82±0.81),차이유통계학의의(P＜0.01);억제VEGF-C적라서림파관밀도명현저우공백대조조(2.62±0.41비5.80±0.81),차이유통계학의의(P＜0.01).동양지,과표체VEGF-C적라서혈관밀도명현고우공백대조조(2.68±0.23비2.01±0.15),차이유통계학의의(P＜0.05);억제VEGF-C적라서혈관밀도명현저우공백대조조(2.01±0.15비1.05±0.47),차이유통계학의의(P＜0.05).동시,아갑람염색결과현시과표체VEGF-C시소서장도종류수목명현다우억제VEGF-C표체적소서(13.4±5.2비4.5±1.8;P＜0.01);이차량조지간장도종류대소야유명현차이.결론 VEGF-C가이통과조절림파내피세포진행촉진장암세포적생장.
Objective Vascular endothelial growth factor-C (VEGF-C) plays important roles in regulation of proliferation, invasion and metastasis of colorectal cancer.However, the concrete mechanism is poorly unknown.The aim of this study was to find out whether VEGF-C promotes colorectal cancer growth through interacting with endothelial lymphatic cells.Methods First the lentiviruses overexpressing or inhibiting VEGF-C were constructed.Western blotting was performed to prove the effectiveness of the lentiviruses.Then three types of CT26 CRC cells (VEGF-C overexpression;VEGF-C inhibition and blank control) were injected into the distal posterior rectum of BALB/c-nude mice.After constructing the animal models successfully, immunofluorescence method was employed to detect the distribution of LYVE-1 and CD31 in lymphatic endothelial tissues and vascular endothelial cells respectively.In addition, the number and size of tumors were compared between the VEGF-C overexpression mice and VEGF-C inhibition mice.Results Western blotting showed that the lentiviruses were successfully constructed.The VEGF-C over-expression group had a significantly higher density of lymph-vessel than in the blank control group (9.10 ± 0.50 vs.5.82 ± 0.81;P ＜ 0.01).The VEGF-C down-expression group had a significantly lower density of lymph-vessel than in the blank control group (2.62 ± 0.41 vs.5.80 ±0.81;P ＜0.01).Similarly, the VEGF-C overexpression group had a significantly higher density of blood-vessel than in the blank control group (2.68 ±0.23 vs.2.01 ±0.15;P＜0.05).The VEGF-C down-expression group had a significantly lower density of lymph-vessel than in the blank control group (2.01 ± 0.15 vs.1.05 ± 0.47;P ＜ 0.05).The number of tumors was more in the VEGF-C overexpression group than in the VEGF-C down-regulation group (13.4 ± 5.2 vs.4.5 ± 1.8;P ＜ 0.01).The tumor size was also significant different.Conclusion VEGF-C might promote colorectal cancer growth through regulating endothelial lymphatic cells.