中华临床营养杂志
中華臨床營養雜誌
중화림상영양잡지
CHINESE JOURNAL OF CLINICAL NUTRITION
2010年
2期
101-105,插1
,共6页
程璐%林岩%曹鹏%蒋苏豫%李苏宜
程璐%林巖%曹鵬%蔣囌豫%李囌宜
정로%림암%조붕%장소예%리소의
重组人生长激素%生长激素受体%血管内皮生长因子%胃癌
重組人生長激素%生長激素受體%血管內皮生長因子%胃癌
중조인생장격소%생장격소수체%혈관내피생장인자%위암
Recombinant human growth hormone%Growth hormone receptor%Vascular endothelial growth factor%Gastric carcinoma
目的 研究重组人生长激素(rhGH)对生长激素受体(GHR)不同表达状态裸鼠人胃癌移植瘤生长及血管内皮生长因子(VEGF)表达的影响.方法 采用免疫细胞化学染色法筛选出GHR阳性和阴性表达的细胞株各1株,分别接种于24只裸鼠皮下.将两种细胞接种裸鼠均随机分为对照组(0.9%NaCl,0.2 ml/d)、低剂量rhGH组(0.5 U·kg-1·d-1,0.2 ml/d)和高剂量rhGH组(2.5 U·kg-1·d-1,0.2 ml/d)3组,每组8只,各组均连续给药14 d,观察并记录裸鼠体重和肿瘤体积变化;采用酶联免疫吸附法测定各组裸鼠血清VEGF含量,免疫组织化学方法检测胃癌组织中VEGF蛋白表达,RT-PCR方法检测胃癌组织VEGF mRNA水平变化.结果 筛选出GHR阳性表达的人胃癌细胞株SGC-7901和阴性表达的MKN-45.对于GHR+SGC-7901接种裸鼠,rhGH给药组皮下移植瘤体积较对照组增大(P＜0.05),且高剂量rhGH组促增长效应最为显著(P＜0.05),3组间体重差异无统计学意义(P＞0.05);高剂量rhGH组的血清VEGF浓度为(252.94±15.32)ng/L,明显高于对照组的(49.94±5.73)ng/L和低剂量rhGH组的(167.60±9.54)ng/L(P＜0.05);对照组VEGF表达为中度阳性,rhGH给药组呈强阳性;高剂量rhGH组肿瘤组织中VEGF mRNA相对表达量为0.6470±0.0447,明显高于对照组的0.3230±0.0258和低剂量rhGH组的0.4120±0.0351(P＜0.05).对于GHR-MKN-45接种裸鼠,rhGH给药组体重明显大于对照组(P＜0.05);肿瘤体积大小、血清VEGF水平、肿瘤组织VEGF蛋白及mRNA表达,3组间差异均无统计学意义(P＞0.05).结论 rhGH能促进GHR阳性表达的SGC-7901移植瘤生长,并促进VEGF表达增高;对于GHR阴性的MKN-45移植瘤,则没有表现出明显的促肿瘤生长及促VEGF表达效应.GHR存在可能是rhGH影响VEGF分泌的关键靶点.
目的 研究重組人生長激素(rhGH)對生長激素受體(GHR)不同錶達狀態裸鼠人胃癌移植瘤生長及血管內皮生長因子(VEGF)錶達的影響.方法 採用免疫細胞化學染色法篩選齣GHR暘性和陰性錶達的細胞株各1株,分彆接種于24隻裸鼠皮下.將兩種細胞接種裸鼠均隨機分為對照組(0.9%NaCl,0.2 ml/d)、低劑量rhGH組(0.5 U·kg-1·d-1,0.2 ml/d)和高劑量rhGH組(2.5 U·kg-1·d-1,0.2 ml/d)3組,每組8隻,各組均連續給藥14 d,觀察併記錄裸鼠體重和腫瘤體積變化;採用酶聯免疫吸附法測定各組裸鼠血清VEGF含量,免疫組織化學方法檢測胃癌組織中VEGF蛋白錶達,RT-PCR方法檢測胃癌組織VEGF mRNA水平變化.結果 篩選齣GHR暘性錶達的人胃癌細胞株SGC-7901和陰性錶達的MKN-45.對于GHR+SGC-7901接種裸鼠,rhGH給藥組皮下移植瘤體積較對照組增大(P＜0.05),且高劑量rhGH組促增長效應最為顯著(P＜0.05),3組間體重差異無統計學意義(P＞0.05);高劑量rhGH組的血清VEGF濃度為(252.94±15.32)ng/L,明顯高于對照組的(49.94±5.73)ng/L和低劑量rhGH組的(167.60±9.54)ng/L(P＜0.05);對照組VEGF錶達為中度暘性,rhGH給藥組呈彊暘性;高劑量rhGH組腫瘤組織中VEGF mRNA相對錶達量為0.6470±0.0447,明顯高于對照組的0.3230±0.0258和低劑量rhGH組的0.4120±0.0351(P＜0.05).對于GHR-MKN-45接種裸鼠,rhGH給藥組體重明顯大于對照組(P＜0.05);腫瘤體積大小、血清VEGF水平、腫瘤組織VEGF蛋白及mRNA錶達,3組間差異均無統計學意義(P＞0.05).結論 rhGH能促進GHR暘性錶達的SGC-7901移植瘤生長,併促進VEGF錶達增高;對于GHR陰性的MKN-45移植瘤,則沒有錶現齣明顯的促腫瘤生長及促VEGF錶達效應.GHR存在可能是rhGH影響VEGF分泌的關鍵靶點.
목적 연구중조인생장격소(rhGH)대생장격소수체(GHR)불동표체상태라서인위암이식류생장급혈관내피생장인자(VEGF)표체적영향.방법 채용면역세포화학염색법사선출GHR양성화음성표체적세포주각1주,분별접충우24지라서피하.장량충세포접충라서균수궤분위대조조(0.9%NaCl,0.2 ml/d)、저제량rhGH조(0.5 U·kg-1·d-1,0.2 ml/d)화고제량rhGH조(2.5 U·kg-1·d-1,0.2 ml/d)3조,매조8지,각조균련속급약14 d,관찰병기록라서체중화종류체적변화;채용매련면역흡부법측정각조라서혈청VEGF함량,면역조직화학방법검측위암조직중VEGF단백표체,RT-PCR방법검측위암조직VEGF mRNA수평변화.결과 사선출GHR양성표체적인위암세포주SGC-7901화음성표체적MKN-45.대우GHR+SGC-7901접충라서,rhGH급약조피하이식류체적교대조조증대(P＜0.05),차고제량rhGH조촉증장효응최위현저(P＜0.05),3조간체중차이무통계학의의(P＞0.05);고제량rhGH조적혈청VEGF농도위(252.94±15.32)ng/L,명현고우대조조적(49.94±5.73)ng/L화저제량rhGH조적(167.60±9.54)ng/L(P＜0.05);대조조VEGF표체위중도양성,rhGH급약조정강양성;고제량rhGH조종류조직중VEGF mRNA상대표체량위0.6470±0.0447,명현고우대조조적0.3230±0.0258화저제량rhGH조적0.4120±0.0351(P＜0.05).대우GHR-MKN-45접충라서,rhGH급약조체중명현대우대조조(P＜0.05);종류체적대소、혈청VEGF수평、종류조직VEGF단백급mRNA표체,3조간차이균무통계학의의(P＞0.05).결론 rhGH능촉진GHR양성표체적SGC-7901이식류생장,병촉진VEGF표체증고;대우GHR음성적MKN-45이식류,칙몰유표현출명현적촉종류생장급촉VEGF표체효응.GHR존재가능시rhGH영향VEGF분비적관건파점.
Objective To investigate the effects of recombinant human growth hormone (rhGH) on tumor growth and vascular endothelial growth factor (VEGF) expression of human gastric carcinoma xenografts in nude mice with different expressions of growth hormone receptor (GHR). Methods Immunocytochemical method was used to pick out one GHR-positive and one GHR-negative cell line. Then the cells were subcutaneously injected into 24 nude mice separately. The nude mice bearing two different kinds of human gastric caicinoma were equalges of body weight and tumor volume of nude mice were recorded. Serum concentrations of VEGF in peripheral blood were analyzed by ELISA. VEGF protein and mRNA expression in tumor tissue were detected by immunohistochemistry and RT-PCR, respectively. Results We chose SGC-7901 as GHR positive group, and MKN-45 as the negative one. For nude mice bearing GHR + SGC-7901 xenografts, the tumor volumes were significantly larger in rhGH groups than in control group (P ＜ 0.05), and the high-dose rhGH group revealed greater effect (P ＜ 0. 05).Body weights were not significantly different among three groups (P ＞ 0. 05). Serum VEGF concentration was (252.94 ± 15.32) ng/L in the high-dose rhGH group, which was significantly higher than that in control group [(49.94 ± 5.73) ng/L] and low-dose rhGH group [(167.60 ± 9.54) ng/L] (P ＜ 0.05). Moderate positive staining with VEGF was observed in the control group, while VEGF staining was strong in rhGH administration groups. The relative expression of VEGF mRNA for the high-dose rhGH group was 0. 6470 ± 0. 0447, which was significantly higher than that in control group (0. 3230 ± 0. 0258)and low-dose rhGH group (0. 412 ± 0. 0351)(P ＜ 0.05). While for nude mice bearing GHR-MKN-45 xenografts, the body weights of the rhGH-administrated groups were significantly higher than that of control group (P ＜ 0.05), while tumor growth, serum VEGF concentration, and the expressions of VEGF mRNA and protein in tumor tissue were not significantly different (P ＞ 0.05).Conclusions rhGH can promote tumor growth and increase the expression of VEGF in the GHR-highly-expressed SGC-7901 xenograft tumor model. However, such effects do not exist in GHR-negatively-expressed MKN-45 xenograft tumor model. The existence of GHR may be a key target where rhGH influences the secretion of VEGF.
