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Experimental method for promoting growth and metastasis of colon cancer cells by lentivirus overexpression vector mediated E6-AP

A colon cancer cell, E6-AP technology, applied in the field of lentiviral overexpression vector-mediated E6-AP to promote colon cancer cell growth and metastasis, to achieve the effect of enhancing growth and inhibiting apoptosis

Inactive Publication Date: 2018-09-25
江西中医药大学第二附属医院
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Problems solved by technology

At the same time, E6-AP is a key protein for normal gonadal development and sexual maturation in the body, and is expressed in various organs in the body, such as testis, prostate, ovary, uterus, breast, brain tissue, stomach, etc.; The mechanism in the process of malignant transformation of cancer has not been reported

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  • Experimental method for promoting growth and metastasis of colon cancer cells by lentivirus overexpression vector mediated E6-AP
  • Experimental method for promoting growth and metastasis of colon cancer cells by lentivirus overexpression vector mediated E6-AP
  • Experimental method for promoting growth and metastasis of colon cancer cells by lentivirus overexpression vector mediated E6-AP

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[0029] The technical solutions of the present invention will be further described in detail below in conjunction with specific embodiments.

[0030] 1. E6AP expression in different colon cancer cell lines

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Abstract

The invention discloses an experimental method for promoting growth and metastasis of colon cancer cells by lentivirus overexpression vector mediated E6-AP. The experimental method comprises the following steps: detecting the expression of E6-AP in different colon cancer cell lines, and determining low expression E6-AP of HCT-116 and HCT-8 colon cancer cells; co-transfecting human embryonic kidney293FT cells with over-expression PCDH-pCDNA3.0-FLAG-E6-AP plasmid and lentiviral packaging helper plasmids to obtain stable expression cells of lentiviral mediated overexpression of E6-AP, and detecting the overexpression of E6-AP through real-time PCR and Western blot detection; and detecting the influence of overexpression of E6-AP on the growth, migration and invasion of colon cancer cells through CCK-8, scratch healing and Transwell TM experiments. The experimental method provides a new target for colon cancer targeted therapy.

Description

technical field [0001] The invention belongs to the field of medicine and biotechnology, and in particular relates to an experimental method for promoting the growth and metastasis of colon cancer cells through E6-AP mediated by a lentiviral overexpression vector. Background technique [0002] Human papillomavirus E6-associated protein (E6AP) is a cell-encoded protein with a relative molecular mass of 100×10 3 ubiquitin protein ligase. Huibregtse et al believe that E6-AP regulates the interaction between the proto-oncoprotein E6 encoded by high-risk human papillomavirus (HPV) 16 and p53, leading to the occurrence of cervical cancer; the study found that E6-AP is the androgen receptor ( AR) co-regulator, enhances the transcriptional activity of various steroid receptors in the body, and has important biological functions for the occurrence and development of prostate cancer and breast cancer. At the same time, E6-AP is a key protein for normal gonadal development and sexual...

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Application Information

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IPC IPC(8): C12N5/10C12N15/867C12Q1/6851C12Q1/06
CPCC12N9/93C12N15/86C12N2740/15043C12Q1/6851C12Y603/02019G01N33/5094C12Q2531/113C12Q2563/107C12Q2545/101
Inventor 熊加秀甘淳左彬荣张奇志晁丹
Owner 江西中医药大学第二附属医院
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