Mouse pancreatic cancer cell line and construction method of homotransplant tumor model

A technology for pancreatic cancer cells and transplanted tumors, which is applied in the field of tumor biology and can solve the problems of poor replication ability of oncolytic herpes simplex virus and inability to reflect anti-tumor functions.

CN110241088AInactive Publication Date: 2019-09-17NANKAI UNIV
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Patent Information

Authority / Receiving Office
CN · China
Current Assignee / Owner
Publication Date
2019-09-17
Estimated Expiration
Not applicable · inactive patent

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Abstract

The invention provides a mouse pancreatic cancer cell line and a construction method of a homotransplant tumor model. The mouse pancreatic cancer cell line Pan02_HR is constructed by a method comprising the following steps of importing a human-source HSV-1 acceptor in a mouse-source pancreatic cancer cell line Pan02 to obtain a mouse pancreatic cancer cell line Pan02_HR stably expressing an HSV-1 acceptor. Through the adoption of the mouse pancreatic cancer cell line Pan02_HR disclosed by the invention, an oncolytic herpes simplex virus can effectively infect and kill the mouse pancreatic cancer cell line, besides, the Pan02_HR cell can form tumors in a mouse body having a complete immune system, and the mouse model can provide an animal model for in vivo assessment of the oncolytic herpes simplex virus and activation of an immune system for treating pancreatic cancer.
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Description

technical field

[0001] The invention belongs to the field of tumor biology, in particular to a mouse pancreatic cancer cell line and a method for constructing a homologous transplantation tumor model. Background technique

[0002] Pancreatic ductal adenocarcinoma (Pancreatic ductal adenocarcinoma, PDAC) is the most common type of pancreatic cancer, accounting for 85% of the total pancreatic cancer, the incidence of pancreatic cancer is very low, less than 3% worldwide, but it has a very high fatality rate. Because of the inconspicuous early symptoms, most patients are diagnosed at an advanced stage. Only 10-20% of patients meet the conditions for surgical resection, and the prognosis is very poor. The postoperative survival period is only 17-20 months. Epidemiological data in my country show that PDAC is currently the sixth cause of cancer-related death in my country, and its incidence has been on the rise in recent years. Although a lot of progress has been made in the ba...

Examples

Embodiment 1

[0028] 1. Construction of mouse pancreatic cancer cell line Pan02_HVEM

[0029] A mouse pancreatic cancer cell line Pan02_HVEM, which is constructed by a method comprising the following steps:

[0030] (1) Construction of pCDH-HVEM-flag-P2A-copGFP lentiviral core vector:

[0031] The HVEM gene was purchased from Beijing Yiqiao Shenzhou Technology Co., Ltd., and the pCDH-MCS-EF1-Puromycin plasmid was purchased from Addgene. The PCR product of flag-P2A-copGFP with BamHI and NotⅠ double restriction sites at both ends was obtained by PCR. The product and the pCDH-MCS-EF1-Puromycin plasmid were double-digested with BamHI and NotⅠ, ligated with T4 ligase, transformed and coated with LB solid plates to obtain single clone colonies, single colonies were picked for small plasmid extraction, and Sanger sequencing confirmed Correct, obtain the pCDH-MCS-flag-P2A-copGFP plasmid;

[0032] The HVEM PCR product with EcoRI and BamHI double restriction sites at both ends was obtained by PCR, ...

Embodiment 2

[0059] 1. Construction of mouse pancreatic cancer cell line Pan02_Nectin-1

[0060] A mouse pancreatic cancer cell line Pan02_Nectin-1, which is constructed by a method comprising the following steps:

[0061] (1) Construction of pCDH-Nectin-1-flag-P2A-copGFP lentiviral core vector:

[0062] The Nectin-1 gene was purchased from Beijing Yiqiao Shenzhou Technology Co., Ltd., and the pCDH-MCS-EF1-Puromycin plasmid was purchased from Addgene. The flag-P2A-copGFP PCR product with BamHI and NotⅠ double restriction sites at both ends was obtained by PCR. The PCR product and the pCDH-MCS-EF1-Puromycin plasmid were digested with BamHI and NotⅠ respectively, ligated with T4 ligase, transformed and coated with LB solid plates to obtain single clone colonies, single colonies were picked for small plasmid extraction, Sanger The sequencing confirmed that it was correct, and the pCDH-MCS-flag-P2A-copGFP plasmid was obtained;

[0063] The Nectin-1 PCR product with BamHI and XbaI double rest...