LAMβ1基因下调VEGF表达抑制人胆管癌裸鼠移植瘤生长
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摘要
目的探讨LAMβ1基因对人胆管癌细胞QBC939裸鼠皮下移植瘤生长的影响。方法构建重组慢病毒LAMβ1过表达载体;将24只裸鼠按随机数字表分为3组,分别为实验组(转染LV-LAMβ1)、阴性对照组(空载慢病毒)以及空白对照组,每组8只。制备密度为1×10~7/ml胆管癌QBC939细胞悬液,取0.2ml分别注入裸鼠皮下,每3d测量瘤体大小,4周后处死裸鼠取出裸鼠皮下瘤体。采用RT-PCR、免疫组化检测瘤组织LAMβ1及VEGF表达。结果成功构建过表达慢病毒LAMβ1载体;与阴性对照组和空白对照组比较,实验组裸鼠皮下移植瘤生长受到抑制,重量与体积明显减小(F_(体积)=18.04,P<0.01;F_(重量)=42.87,P<0.01);实验组裸鼠瘤组织中LAMβ1 mRNA和蛋白水平较阴性对照组及空白对照组增高,转移因子VEGF mRNA和蛋白水平下降(P<0.05)。结论 LAMβ1基因过表达通过下调转移因子VEGF表达抑制胆管癌裸鼠皮下移植瘤的生长。
Objective To explore the effects of LAMB1 gene on growth of xenografts in nude mice. Methods The recombinant chronic virus LAMB1 overexpression vector was constucted. 24 female nude mice were randomly divided into exprimental group(LV-LAMB1), negative control group and blank control group. Colorectal carcinoma QBC939 cell suspension(0.2 ml, 1×10~7/ml) was injected into nude mice. Tumor volume and weight were measured every 3 d. Nude mice were sacrificed 4 weeks later and tumor samples removed. LAMB1 and VEGF expression in tumor samples were detected by RT-PCR and immunohistochemistry. Results Recombinant chronic virus LAMB1 overexpression vector was successfully constructed. Compared with negative control group and blank control group, the growth rate of subcutaneous xenograft significantly decreased in experimental group. The tumor's size and weight of experimental group were significantly lesser than those of the two others group(Volume, F=18.04, P<0.01; Weight, F=42.87, P<0.01). The expression of LAMB1 were higher than that in the two other groups(P<0.05). The expression of VEGF was higher than that in the experimental groups(P<0.05). Conclusions LV-LAMB1 can inhibit the growth of biliary duct cancer. LAMB1 in QBC939 can inhibit the expression of VEGF, which was related to biliary duct cancer metastasis.
Objective To explore the effects of LAMB1 gene on growth of xenografts in nude mice. Methods The recombinant chronic virus LAMB1 overexpression vector was constucted. 24 female nude mice were randomly divided into exprimental group(LV-LAMB1), negative control group and blank control group. Colorectal carcinoma QBC939 cell suspension(0.2 ml, 1×10~7/ml) was injected into nude mice. Tumor volume and weight were measured every 3 d. Nude mice were sacrificed 4 weeks later and tumor samples removed. LAMB1 and VEGF expression in tumor samples were detected by RT-PCR and immunohistochemistry. Results Recombinant chronic virus LAMB1 overexpression vector was successfully constructed. Compared with negative control group and blank control group, the growth rate of subcutaneous xenograft significantly decreased in experimental group. The tumor's size and weight of experimental group were significantly lesser than those of the two others group(Volume, F=18.04, P<0.01; Weight, F=42.87, P<0.01). The expression of LAMB1 were higher than that in the two other groups(P<0.05). The expression of VEGF was higher than that in the experimental groups(P<0.05). Conclusions LV-LAMB1 can inhibit the growth of biliary duct cancer. LAMB1 in QBC939 can inhibit the expression of VEGF, which was related to biliary duct cancer metastasis.
引文
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[2] Liu T,Yang H,Fan W,et al.Mechanisms of MAFG Dysregulation in Cholestatic Liver Injury and Development of Liver Cancer[J].Gastroenterology,2018,155(2):557-571.
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[4] Ogura T,Miyano A,Nishioka N,et al.Recanalization for Tight Bile Duct-Jejunum Anastomosis Stricture Using Peroral Transliminal Cholangioscopy (with Video)[J].Dig Dis,2018,28(3):1-4.
[5] Satoh K.Strong carcinogenic stress response induction of preneoplastic cells positive for GST-P in the rat liver:Physiological mechanism for initiation[J].Life Sci,2018,200:42-48.
[6] Wang D,Korhonen PK,Gasser RB,et al.Improved genomic resources and new bioinformatic workflow for the carcinogenic parasite Clonorchis sinensis:Biotechnological implications[J].Biotechnol Adv,2018,36(4):894-904.
[7] Lin YK,Fang Z,Jiang TY,et al.Combination of Kras activation and PTEN deletion contributes to murine hepatopancreatic ductal malignancy[J].Cancer Lett,2018,421:161-169.
[8] Wagner JUG,Chavakis E,Rogg EM,et al.Switch in Laminin β2 to Laminin β1 Isoforms During Aging Controls Endothelial Cell Functions-Brief Report[J].Arterioscler Thromb Vasc Biol,2018,38(5):1170-1177.
[9] Wardell CP,Fujita M,Yamada T,et al.Genomic characterization of biliary tract cancers identifies driver genes and predisposing mutations[J].J Hepatol,2018,68(5):959-969.
[10] Ji X,Wu B,Han R,et al.The association of LAMB1 polymorphism and expression changes with the risk of coal workers' pneumoconiosis[J].Environ Toxicol,2017,32(9):2182-2190.
[11] Xu Y,Yao Y,Zhong X,et al.Downregulated circular RNA hsa_circ_0001649 regulates proliferation,migration and invasion in cholangiocarcinoma cells[J].Biochem Biophys Res Commun,2018,496(2):455-461.
[12] Herrera-Goepfert R.Postradiation Synovial Sarcoma of the Common Bile Duct:A Previously Unreported Anatomic Site[J].Int J Surg Pathol,2018,26(5):469-474.
[13] Kim SJ,Akita M,Sung YN,et al.MDM2 Amplification in Intrahepatic Cholangiocarcinomas:Its Relationship With Large-Duct Type Morphology and Uncommon KRAS Mutations[J].Am J Surg Pathol,2018,42(4):512-521.
[14] Suzumura K,Hatano E,Tada M,et al.Difficulty Achieving a Preoperative Diagnosis of IgG4-Related Sclerosing Cholangitis[J].Case Rep Gastroenterol,2018,12(2):425-431.
[15] Andersson ER,Chivukula IV,Hankeova S,et al.Mouse Model of Alagille Syndrome and Mechanisms of Jagged1 Missense Mutations[J].Gastroenterology,2018,154(4):1080-1095.
[16] Broutier L,Mastrogiovanni G,Verstegen MM,et al.Human primary liver cancer-derived organoid cultures for disease modeling and drug screening[J].Nat Med,2017,23(12):1424-1435.
[17] Amblard I,Mercier F,Bartlett DL,et al.Cytoreductive surgery and HIPEC improve survival compared to palliative chemotherapy for biliary carcinoma with peritoneal metastasis:A multi-institutional cohort from PSOGI and BIG RENAPE groups[J].Eur J Surg Oncol,2018,44(9):1378-1383.
[18] Matsumoto T,Takai A,Eso Y,et al.Proliferating EpCAM-Positive Ductal Cells in the Inflamed Liver Give Rise to Hepatocellular Carcinoma[J].Cancer Res,2017,77(22):6131-6143.
[19] Guragain D,Seubwai W,Kobayashi D,et al.Artesunate and chloroquine induce cytotoxic activity on cholangiocarcinoma cells via different cell death mechanisms[J].Cell Mol Biol,2018,64(10):113-118.
[20] Thongsom S,Suginta W,Lee KJ,et al.Piperlongumine induces G2/M phase arrest and apoptosis in cholangiocarcinoma cells through the ROS-JNK-ERK signaling pathway[J].Apoptosis,2017,22(11):1473-1484.