Application of novel oncolytic virus in preparation of medicine for treating colorectal cancer
An oncolytic virus, colorectal cancer technology, applied in the fields of biotechnology and biomedicine, can solve the problems of lack of specificity of infecting tumors, difficult to guarantee safety, unstable genetic characteristics of RNA oncolytic virus, etc., and achieve large clinical application. Promising, easy to handle, effect of significant oncolytic properties
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Embodiment 1
[0060] Embodiment 1: the killing effect detection of EV71 in vitro
[0061] The killing effect detection of EV71 on colorectal cancer cells, the steps are:
[0062] 1. Experimental materials:
[0063] 1.1 Cells and viruses:
[0064] DLD1, HCT116, LoVo, HT29, SW48 colorectal cancer cell lines were obtained from ATCC Company in the United States. Enterovirus 71 (EV71) strain (Xiangyang-Hubei-09) was derived from EV71-infected brain tissue in Xiangyang City, Hubei Province, China, and was previously isolated from the State Key Laboratory of Virology, Wuhan University (GenBank: JN230523.1). species expanded in RD cells.
[0065] 1.2 Reagents:
[0066] DMEM medium and fetal bovine serum (FBS) were purchased from GIBCO; CCK8 (Cell Counting Kit-8) detection kit was purchased from Sangon Bioengineering (Shanghai) Co., Ltd.
[0067] 1.3 Experimental equipment:
[0068] The microplate reader is a product of Thermo Company; the cell culture box is a product of Thermo Company.
[0...
Embodiment 2
[0082] Embodiment 2: In vitro EV71 detects the killing effect of normal intestinal cells
[0083] The present invention provides a kind of application of human enterovirus EV71 in human colorectal cancer, and its steps are:
[0084] 1. Experimental materials
[0085] 1.1 Cells and viruses
[0086] Human primary intestinal epithelial cells FHC and HCoEpic were purchased from ATCC, USA. EV71 (Human enterovirus71strain Xiangyang-Hubei-09) was isolated from the State Key Laboratory of Virology of Wuhan University (GenBank: JN230523.1), and the virus stocks were amplified in RD cells.
[0087] 1.2 Reagents
[0088] DMEM medium and FBS were purchased from GIBCO; CCK8 (Cell Counting Kit-8) detection kit was purchased from Sangon Bioengineering (Shanghai) Co., Ltd.
[0089] 1.3 Experimental Instruments
[0090] The microplate reader is a product of Thermo Company; the cell culture box is a product of Thermo Company.
[0091] 2. Experimental methods and results
[0092] 2.1 Dete...
Embodiment 3
[0101] Embodiment 3: the detection of the oncolytic effect of EV71 in vivo
[0102] The application of a kind of human enterovirus EV71 of the present invention in human colorectal cancer, its steps are:
[0103] 1. Experimental materials:
[0104] 1.1 Cells, viruses, nude mice:
[0105] HT29 cells were purchased from ATCC, USA. Nude mice (5 weeks old, male) were purchased from the Experimental Animal Center of Guangdong Province, China. EV71 (Human enterovirus 71strain Xiangyang-Hubei-09) was isolated from the State Key Laboratory of Virology of Wuhan University (GenBank: JN230523.1), and the virus stock was amplified in RD cells.
[0106] 1.2 Reagents:
[0107] DMEM medium, FBS and PBS buffer solution (pH=7.4) were purchased from GIBCO Company.
[0108] 1.3 Experimental equipment:
[0109] The cell incubator is a product of Thermo Company.
[0110] 2. Experimental methods and results:
[0111] 2.1 Construction of xenograft tumor transplantation model in nude mice:
...
PUM
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Abstract
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