Coxsackievirus for treating tumor
A coxsackie virus, tumor technology, applied in the field of virus and tumor treatment
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Embodiment 1
[0262] Example 1: Acquisition and preparation of CVB3 and its modified forms
[0263] 1.1 Isolation of enterovirus CVB3 from clinical samples of patients
[0264] (1) The patients' throat swabs and anal swabs are from China, Xiamen Center for Disease Control and Prevention; African green monkey kidney cells (Vero cells; Number: CCL-81 TM ) is preserved by China, Xiamen University, National Engineering Technology Research Center for Diagnostic Reagents and Vaccines of Infectious Diseases, and cultured in MEM medium supplemented with 10% fetal bovine serum, glutamine, penicillin and streptomycin.
[0265] (2) Specimen processing: Fully stir the patient's throat swab and anal swab in the specimen preservation solution to wash off the virus and virus-containing cells attached to the swab, and then store the specimen preservation solution at 4°C 4000rpm high-speed centrifugation for 30min;
[0266] (3) Inoculation and observation:
[0267] A. Spread Vero cells on a 24-well p...
Embodiment 2
[0284] Example 2: In vitro anti-tumor experiments of CVB3 and its modified forms
[0285] 2.1 Viruses and cell lines used
[0286] (1) Viruses: CVB3-WT (SEQ ID NO: 12), CVB3-HRV2 (SEQ ID NO: 13), CVB3-miR133&206T (SEQ ID NO: 14), CVB3- GM-CSF (SEQ ID NO: 15) and CVB3-Anti-PD-1 (SEQ ID NO: 16).
[0287] (2) Cell line: human rhabdomyosarcoma cell RD ( Number: CCL-136 TM ); human colorectal cancer cell line SW1116 ( Number: CCL-233 TM ), SW480 ( Number: CCL-228 TM ) and HT-29 ( Number:HTB-38 TM ); human gastric cancer cell line AGS ( Number: CRL-1739 TM ), SGC7901 (CCTCC deposit number: GDC150), BGC823 (CCTCC deposit number: GDC151) and NCI-N87 ( Number: CRL-5822 TM ); human esophageal carcinoma cell line TE-1 (purchased from the Cell Resource Center, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, number: 3131C0001000700089); human small cell lung cancer cell line DMS114 ( Number: CRL-2066 TM ); human non-small cell lung cancer ce...
Embodiment 3
[0329] Example 3: In vivo anti-tumor experiments of CVB3 and its modified forms
[0330] 3.1 Viruses, cell lines and experimental animals
[0331] (1) Viruses: CVB3-WT (SEQ ID NO: 12), CVB3-HRV2 (SEQ ID NO: 13), CVB3-miR133&206T (SEQ ID NO: 14), CVB3 provided in Example 1 were used in this example - GM-CSF (SEQ ID NO: 15) and CVB3-Anti-PD-1 (SEQ ID NO: 16). Refer to Examples 2.2 and 2.3 for the methods of virus cultivation and virus titer determination, respectively.
[0332] (2) Cell lines: human breast cancer cell line BcaP37 (CCTCC deposit number: GDC206), human non-small cell lung cancer cell line A549 ( Number: CCL-185 TM ), human Burkitt's lymphoma cell line Raji ( Number: CCL-86 TM ), human endometrial cancer cell line Ishikawa (ECACC No.99040201), human cervical cancer cell line Hela ( Number: CCL-2 TM ) and human glioma cell line GBM (primary tumor cell line isolated from patient tumor tissue). The above cells except Raji used RPMI-1640 medium, and other ...
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