Construction and expression of prokaryotic expression vector of vascular endothelial growth factor
PANG Lijun1 LIU Daojie2 LIN Minghua1 LIU Kai1 LIU Xiaoni1 CHEN Dexi1
1.Beijing You′an Hospital, Capital Medical University Beijing Institute of Hepatology, Beijing 100069, China;
2.Department of Clinical Laboratory, Haidian Maternal and Child Health Hospital, Beijing 100080, China
Abstract：Objective To construct a prokaryotic expression vector containing vascular endothelial growth factor (VEGF) and induce to express protein, it may lay a foundation for the follow-up study of VEGF. Methods Amplified The target fragment was amplified by reverse transcription polymerase chain reaction (RT-PCR) and the DNA fragment was cloned into pET-30a(+) vector with histidine tag. The recombinant plasmid was identified by polymerase chain reaction (PCR) and sequencing. The expression of recombinant plasmid was induced by isopropylthio-β-D-galactoside (IPTG) and identified by Western blot. Results The right size of the fragment was amplified from HepG2 cells. The recombinant plasmid pET-30a-VEGF was confirmed by PCR and DNA sequencing. Coomassie brilliant blue staining and Western blot results showed that the molecular weight of the fusion protein induced by IPTG was correct and the band was single. Conclusion The prokaryotic expression plasmid of pET-30a-VEGF is successfully constructed, in addition, it might lay the foundation for development of HCC diagnostic and treatment research.
庞丽君1 刘道洁2 林明华1 刘凯1 刘晓霓1 陈德喜1. 血管内皮生长因子原核表达载体的构建及表达[J]. 中国医药导报, 2020, 17(23): 4-7.
PANG Lijun1 LIU Daojie2 LIN Minghua1 LIU Kai1 LIU Xiaoni1 CHEN Dexi1. Construction and expression of prokaryotic expression vector of vascular endothelial growth factor. 中国医药导报, 2020, 17(23): 4-7.
[1] Yang JD，Hainaut P，Gores GJ，et al. A global view of hepatocellular carcinoma：trends，risk，prevention and management [J]. Nat Rev Gastroenterol Hepatol，2019，16（10）：589-604.
[2] Lee HW，Cho KJ，Park JY. Current Status and Future Direction of Immunotherapy in Hepatocellular Carcinoma：What Do the Data Suggest? [J]. Immune Netw，2020，20（1）：e11.
[3] Akinyemiju T，Abera S，Ahmed M，et al. The Burden of Primary Liver Cancer and Underlying Etiologies From 1990 to 2015 at the Global，Regional，and National Level：Results From the Global Burden of Disease Study 2015 [J]. JAMA Oncol，2017，3（12）：1683-1691.
[4] Kim BH，Park JW. Epidemiology of liver cancer in South Korea [J]. Clin Mol Hepatol，2018，24（1）：1-9.
[5] Lee JS，Cho IR，Lee HW，et al. Conditional Survival Estimates Improve Over Time for Patients with Hepatocellular Carcinoma：An Analysis for Nationwide Korea Cancer Registry Database [J]. Cancer Res Treat，2019，51（4）：1347-1356.
[6] Wang D，Luo L，Chen W，et al. Significance of the vascular endothelial growth factor and the macrophage migration inhibitory factor in the progression of hepatocellular carcinoma [J]. Oncol Rep，2014，31（3）：1199-1204.
[7] Niu G，Chen X. Vascular endothelial growth factor as an anti-angiogenic target for cancer therapy [J]. Curr Drug Targets，2010，11（8）：1000-1017.
[8] Zhang W，Kim R，Quintini C，et al. Prognostic role of plasma vascular endothelial growth factor in patients with hepatocellular carcinoma undergoing liver transplantation [J]. Liver Transpl，2015，21（1）：101-111.
[9] Syed YY. Ramucirumab：A Review in Hepatocellular Carcinoma [J]. Drugs，2020，80（3）：315-322.
[10] Medavaram S，Zhang Y. Emerging therapies in advanced hepatocellular carcinoma [J]. Exp Hematol Oncol，2018， 7：17.
[11] Li TY，Yang Y，Zhou G，et al. Immune suppression in chronic hepatitis B infection associated liver disease：A review [J]. World J Gastroenterol，2019，25（27）：3527-3537.
[12] Villanueva A. Hepatocellular Carcinoma [J]. N Engl J Med，2019，380（15）：1450-1462.
[13] Lou W，Liu J，Gao Y，et al. MicroRNA regulation of liver cancer stem cells [J]. Am J Cancer Res，2018，8（7）：1126-1141.
[14] Kim DW，Talati C，Kim R. Hepatocellular carcinoma（HCC）：beyond sorafenib-chemotherapy [J]. J Gastrointest Oncol，2017，8（2）：256-265.
[15] Kirstein MM，Wirth TC. Multimodal treatment of hepatocellular carcinoma [J]. Internist：Berl，2020，61（2）：164-169.
[16] Zhu AX，Duda DG，Sahani DV，et al. HCC and angiogenesis：possible targets and future directions [J]. Nat Rev Clin Oncol，2011，8（5）：292-301.
[17] Bangaru S，Marrero JA，Singal AG. Review article：new therapeutic interventions for advanced hepatocellular carcinoma [J]. Aliment Pharmacol Ther，2020，51（1）：78-89.
[18] Rossetto A，Re VD，Steffan A，et al. Carcinogenesis and Metastasis in Liver：Cell Physiological Basis [J]. Cancers：Basel，2019，11（11）：1731.
[19] Terme M，Colussi O，Marcheteau E，et al. Modulation of immunity by antiangiogenic molecules in cancer [J]. Clin Dev Immunol，2012，2012：492920.
[20] Rivera LB，Meyronet D，Hervieu V，et al. Intratumoral myeloid cells regulate responsiveness and resistance to antiangiogenic therapy [J]. Cell Rep，2015，11（4）：577-591.
[21] Gabrilovich DI，Ostrand-Rosenberg S，Bronte V. Coordinated regulation of myeloid cells by tumours [J]. Nat Rev Immunol，2012，12（4）：253-268.
[22] Voron T，Colussi O，Marcheteau E，et al. VEGF-A modulates expression of inhibitory checkpoints on CD8+ T cells in tumors [J]. J Exp Med，2015，212（2）：139-148.
[23] Kudo M. Scientific Rationale for Combined Immunotherapy with PD-1/PD-L1 Antibodies and VEGF Inhibitors in Advanced Hepatocellular Carcinoma [J]. Cancers：Basel，2020，12（5）：1089.
[24] Hato T， Zhu AX， Duda DG. Rationally combining anti-VEGF therapy with checkpoint inhibitors in hepatocellular carcinoma [J]. Immunotherapy，2016，8（3）：299-313.
[25] Cheng AL，Hsu C，Chan SL，et al. Challenges of combination therapy with immune checkpoint inhibitors for hepatocellular carcinoma [J]. J Hepatol，2020，72（2）：307-319.
2020, Vol. 17 
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