冷冻消融联合GM-CSF治疗对肾癌小鼠血管生成及细胞增殖的影响
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摘要
目的探讨冷冻消融联合粒细胞-巨噬细胞集落刺激因子(GM-CSF)治疗对肾癌小鼠血管生成及细胞增殖的影响。方法取40只健康级别为SPF级的BALB小鼠,肾癌细胞株786-0建立荷肾癌小鼠模型,将模型建立成功的荷肾癌小鼠按随机数字表法分A、B、C、D四组,每组10只,A组不做任何处理,B组给予GM-CSF,C组给予冷冻消融处理,D组给予冷冻消融联合GM-CSF治疗,记录小鼠基本生存状态,观察小鼠肿瘤变化情况,绘制瘤体大小折线图,并分别应用HUVEC小管形成实验、MTT法检测各组小鼠血管生成及细胞增殖情况。结果小鼠接种肾癌细胞株786-0后未见明显进食异常现象,有轻度精神萎靡、纳差,接种部位随时间延长而逐渐膨隆,但接种部位表皮未见破溃、出血现象,接种2周时无死亡小鼠,HE染色证实荷肾癌小鼠模型成功建立;A组治疗后1、2、3周时瘤体逐渐增大,B组亦持续增大,但增大幅度较A组小,C组、D组在治疗2周后便可见瘤体缩小,4组小鼠不同时间点的瘤体大小差异有统计学意义(P<0.05);且治疗2周、4周时C、D组小鼠瘤体显著小于A、B组(P<0.05);B、C、D组管腔形成长度、个数均显著低于A组(P<0.05),而D组管腔形成长度、个数显著低于B、C组(P<0.05);B、C、D 3组小鼠细胞治疗24、48、72、96、128h时的细胞增殖抑制率比较差异均有统计学意义(P<0.05),且治疗后72h细胞增殖抑制率最高,至96、128h均有下降,但D组任意时间段的细胞增殖抑制率均显著高于B、C组(P<0.05)。结论冷冻消融联合GM-CSF治疗可显著抑制肾癌小鼠血管生成及细胞增殖,缩小瘤体体积,且安全性良好。
Objective To analyze the effects of cryoablation combined with granulocyte-macrophage colony-stimulating factor(GM-CSF)on angiogenesis and cell proliferation in renal carcinoma mice.Methods 40 BALB mice with the health level at SPF were taken.The renal carcinoma mouse model was established with renal carcinoma cell line 786-0.The mice with renal carcinoma successful establishing the model were randomly divided into A,B,C,and D group.Group A did not execute any treatment,group B was given GM-CSF,group C was given cryoablation treatment and group D was given cryoablation combined with GM-CSF.The basic survival status of the mice was recorded.The tumor changes of the mice were observed.The line chart of tumor size was drawn.The HUVEC tubule formation experiment and MTT were respectively used to detect angiogenesis and cell proliferation in each group.Results After the mice were inoculated with renal carcinoma cell line 786-0,there was no obvious abnormal eating phenomenon with mild energielos,poor appetite.The inoculation site gradually expanded with time.The epidermis of the inoculation site showed no ulceration and bleeding.At 2 weeks after inoculation,there was no death mouse.HE staining confirmed that the mouse model with renal carcinoma was successfully established.In the group A,the tumor gradually increased at 1,2,and 3 weeks after treatment.The tumor in the group A also continued to increase.The increase degree of group A was larger than that of group B.The tumor became small in group C and group D at 2 weeks after treatment.There was statistical significance in the difference of tumor size in the 4 groups at different time points,the tumor size in groups C and D at 2 and 4 weeks after treatment was significantly smaller than that in group A and B(P<0.05).The length and number of lumen formation in group B,C and D were significantly lower than those in group A(P<0.05),while the length and number of lumen formation in group D were significantly lower than those in group B and C(P<0.05).The difference in cell proliferation inhibition rates of the mice in group B,C and D at 24 h,48 h,72 h,96 hand 128 hwas statistically significant the(P<0.05).The inhibition rate of cell proliferation was the highest at 72 hafter treatment,it decreased at 96 hand128 h,the inhibition rate of cell proliferation in group D was significantly higher than that in group B and C at any time(P<0.05).Conclusion Cryoablation combined with GM-CSF therapy can significantly inhibit angiogenesis and cell proliferation in renal carcinoma mice and reduce tumor volume.The safety is good.
Objective To analyze the effects of cryoablation combined with granulocyte-macrophage colony-stimulating factor(GM-CSF)on angiogenesis and cell proliferation in renal carcinoma mice.Methods 40 BALB mice with the health level at SPF were taken.The renal carcinoma mouse model was established with renal carcinoma cell line 786-0.The mice with renal carcinoma successful establishing the model were randomly divided into A,B,C,and D group.Group A did not execute any treatment,group B was given GM-CSF,group C was given cryoablation treatment and group D was given cryoablation combined with GM-CSF.The basic survival status of the mice was recorded.The tumor changes of the mice were observed.The line chart of tumor size was drawn.The HUVEC tubule formation experiment and MTT were respectively used to detect angiogenesis and cell proliferation in each group.Results After the mice were inoculated with renal carcinoma cell line 786-0,there was no obvious abnormal eating phenomenon with mild energielos,poor appetite.The inoculation site gradually expanded with time.The epidermis of the inoculation site showed no ulceration and bleeding.At 2 weeks after inoculation,there was no death mouse.HE staining confirmed that the mouse model with renal carcinoma was successfully established.In the group A,the tumor gradually increased at 1,2,and 3 weeks after treatment.The tumor in the group A also continued to increase.The increase degree of group A was larger than that of group B.The tumor became small in group C and group D at 2 weeks after treatment.There was statistical significance in the difference of tumor size in the 4 groups at different time points,the tumor size in groups C and D at 2 and 4 weeks after treatment was significantly smaller than that in group A and B(P<0.05).The length and number of lumen formation in group B,C and D were significantly lower than those in group A(P<0.05),while the length and number of lumen formation in group D were significantly lower than those in group B and C(P<0.05).The difference in cell proliferation inhibition rates of the mice in group B,C and D at 24 h,48 h,72 h,96 hand 128 hwas statistically significant the(P<0.05).The inhibition rate of cell proliferation was the highest at 72 hafter treatment,it decreased at 96 hand128 h,the inhibition rate of cell proliferation in group D was significantly higher than that in group B and C at any time(P<0.05).Conclusion Cryoablation combined with GM-CSF therapy can significantly inhibit angiogenesis and cell proliferation in renal carcinoma mice and reduce tumor volume.The safety is good.
引文
[1]李汉忠,孙颖浩,魏强,等.中国18家医院2008-2012年肾癌诊治现状分析[J].中华泌尿外科杂志,2014,35(6):406-409.
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[6] Helft,Julie,B ttcher,et al.GM-CSF Mouse Bone MarrowCultures Comprise a Heterogeneous Population of CD11c+MH-CII+Macrophages and Dendritic Cells[J].Immunity,2015,42(6):1197-1211.
[7] Lutz M B,Kukutsch N A,Menges M,et al.Culture of bonemarrow cells in GM-CSF plus high doses of lipopolysaccharidegenerates exclusively immature dendritic cells which induce al-loantigen-specific CD4Tcell anergy in vitro[J].European Jour-nal of Immunology,2015,30(4):1048-1052.
[8]杨书坤,王明.肾癌保留肾单位手术与根治性肾切除手术的诊疗比较[J].第三军医大学学报,2014,36(23):2404-2407.
[9] Weaver M L,Atkinson D,Zemel R.Hepatic cryosurgery intreating colorectal metastases[J].Cancer,2015,76(2):210-214.
[10] NORDIN.Curettage cryosurgery for non melanoma skin cancerof the external ear:excellent 5‐year results[J].British Jour-nal of Dermatology,2015,140(2):291-293.
[11] Abramovits W,Graham G,Har-Shai Y,et al.DermatologicalCryosurgery and Cryotherapy[J].Anticancer Research,2016,36(9):4979.
[12]魏世平,李辉明,陶维雄,等.腹腔镜下冷冻消融治疗肾癌的长期随访报告[J].中国微创外科杂志,2015,15(4):344-346.
[13]井亮亮,苗梓韬,李曼曼,等.融合蛋白hVEGF121/βhCG与mGM-CSF/βhCG联合抗肿瘤作用及其机制[J].中国药科大学学报,2017,48(1):102-109.
[14]冯婷婷,花道金,韩正祥,等.rhGM-CSF和rhG-CSF对A549细胞顺铂化疗敏感性的影响(The effects of rhGM-CSF and rhG-CSF on chemosensitivity of A549cells to cisplatin)[J].肿瘤,2016,34(7):609-615.
[15] Simmons S,Tjoa B M,Elgamal A,et al.GM-CSF as a system-ic adjuvant in a phase II prostate cancer vaccine trial[J].Pros-tate,2015,39(4):291-297.
[16]费夏玮,刘光香,汪维,等.氩氦刀冷冻消融治疗兔VX2肾癌疗效的初步研究[J].中华泌尿外科杂志,2014,35(6):442-446.
[17]尹天泉,张米,何新华.氩氦刀治疗小鼠肺腺癌后瘤周注射树突状细胞对冷冻免疫效应的影响[J].中国实验诊断学,2016,20(10):1631-1634.
[18]牛超,许建婷,徐东升,等.IFN-α联合GM-CSF诱导胃癌患者外周血单个核细胞分化为树突状细胞[J].中国肿瘤生物治疗杂志,2013,20(4):404-408.
[2]黄翼然,张进,陈勇辉,等.“球冠状”肾部分切除术治疗早期肾癌的临床研究[J].中华泌尿外科杂志,2015,36(3):166-171.
[3]燕翔,汪维,常晓峰,等.局麻下经皮冷冻消融治疗高手术风险T1期肾癌的疗效和安全性分析[J].中华泌尿外科杂志,2014,35(6):418-421.
[4]徐斌,宋尚卿,吴震杰,等.肾癌冷冻消融术64例经验总结[J].中华泌尿外科杂志,2018,41(6):422-427.
[5] De L T A,Benson M C,Bagiella E,et al.Cryoablation for clin-ically localized prostate cancer using an argon-based system:complication rates and biochemical recurrence[J].Bju Interna-tional,2015,85(3):281-286.
[6] Helft,Julie,B ttcher,et al.GM-CSF Mouse Bone MarrowCultures Comprise a Heterogeneous Population of CD11c+MH-CII+Macrophages and Dendritic Cells[J].Immunity,2015,42(6):1197-1211.
[7] Lutz M B,Kukutsch N A,Menges M,et al.Culture of bonemarrow cells in GM-CSF plus high doses of lipopolysaccharidegenerates exclusively immature dendritic cells which induce al-loantigen-specific CD4Tcell anergy in vitro[J].European Jour-nal of Immunology,2015,30(4):1048-1052.
[8]杨书坤,王明.肾癌保留肾单位手术与根治性肾切除手术的诊疗比较[J].第三军医大学学报,2014,36(23):2404-2407.
[9] Weaver M L,Atkinson D,Zemel R.Hepatic cryosurgery intreating colorectal metastases[J].Cancer,2015,76(2):210-214.
[10] NORDIN.Curettage cryosurgery for non melanoma skin cancerof the external ear:excellent 5‐year results[J].British Jour-nal of Dermatology,2015,140(2):291-293.
[11] Abramovits W,Graham G,Har-Shai Y,et al.DermatologicalCryosurgery and Cryotherapy[J].Anticancer Research,2016,36(9):4979.
[12]魏世平,李辉明,陶维雄,等.腹腔镜下冷冻消融治疗肾癌的长期随访报告[J].中国微创外科杂志,2015,15(4):344-346.
[13]井亮亮,苗梓韬,李曼曼,等.融合蛋白hVEGF121/βhCG与mGM-CSF/βhCG联合抗肿瘤作用及其机制[J].中国药科大学学报,2017,48(1):102-109.
[14]冯婷婷,花道金,韩正祥,等.rhGM-CSF和rhG-CSF对A549细胞顺铂化疗敏感性的影响(The effects of rhGM-CSF and rhG-CSF on chemosensitivity of A549cells to cisplatin)[J].肿瘤,2016,34(7):609-615.
[15] Simmons S,Tjoa B M,Elgamal A,et al.GM-CSF as a system-ic adjuvant in a phase II prostate cancer vaccine trial[J].Pros-tate,2015,39(4):291-297.
[16]费夏玮,刘光香,汪维,等.氩氦刀冷冻消融治疗兔VX2肾癌疗效的初步研究[J].中华泌尿外科杂志,2014,35(6):442-446.
[17]尹天泉,张米,何新华.氩氦刀治疗小鼠肺腺癌后瘤周注射树突状细胞对冷冻免疫效应的影响[J].中国实验诊断学,2016,20(10):1631-1634.
[18]牛超,许建婷,徐东升,等.IFN-α联合GM-CSF诱导胃癌患者外周血单个核细胞分化为树突状细胞[J].中国肿瘤生物治疗杂志,2013,20(4):404-408.