Method for inhibiting hepatoma carcinoma nude mouse transplantation tumor capacity through silent secretory clusterin

A technology of clustering proteins and inhibiting liver cancer, which is applied in the direction of introducing foreign genetic material using vectors, recombinant DNA technology, animal husbandry, etc., can solve the problems of loss of surgical treatment, lack of effective treatment methods for advanced liver cancer, etc., and achieve simple and efficient methods and delay growth speed effect

Inactive Publication Date: 2017-02-22
AFFILIATED HOSPITAL OF NANTONG UNIV
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  • Application Information

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Problems solved by technology

At present, surgical resection or liver transplantation is still the first choice for treatment, but due to the rapid development of hepatocellular carcinoma, most patients often lose the opportunity for surgical treatment when they are diagnosed, and can only be treated with radiotherapy, chemotherapy and local treatment. There is still no effective treatment for liver cancer

Method used

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  • Method for inhibiting hepatoma carcinoma nude mouse transplantation tumor capacity through silent secretory clusterin
  • Method for inhibiting hepatoma carcinoma nude mouse transplantation tumor capacity through silent secretory clusterin
  • Method for inhibiting hepatoma carcinoma nude mouse transplantation tumor capacity through silent secretory clusterin

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Embodiment 1

[0037] This example is the specific steps of the intervention and detection of the ability of silencing secreted clusterin to inhibit liver cancer xenografts in nude mice of the present invention.

[0038] 1. Cell culture: Human liver cancer cell line HepG2 was cultured in 10% FCS DMEM medium, and the cell line was placed in 5% CO 2 , 37 ℃ incubator, conventional culture.

[0039] 2. Transfect shRNA: design interference sequence according to human secreted clusterin sequence (NCBI: NM_001831.3): shRNA-1, 5'-GTAAGTACGTCAATAAGGA-3' and negative control sequence NC-shRNA, 5'-TTCTCCGAACGTGTCACGT-3 ', and construct the plasmid vector pRNAT-U6.1 / Neo-shRNA1; inoculate HepG2 / ADM cells in a six-well plate, and wait until they grow to 80% of the total well. NC-shRNA was transferred into HepG2 cells, and blank control wells were set; after 12 hours, the original medium was replaced to continue the culture, and after 24 hours, the transfection efficiency was observed by fluorescence micr...

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Abstract

The invention discloses a method for inhibiting hepatoma carcinoma nude mouse transplantation tumor capacity through silent secretory clusterin. The method includes the two steps of interference and detection. The method has the advantages that the method is simple and efficient, and it is clearly shown that the silent secretory clusterin can obviously inhibit the human hepatoma carcinoma nude mouse subcutaneous tumor formation capacity. It is effectively shown that the transcription of the silent secretory clusterin prolongs the incubation period of a human hepatoma carcinoma cell tumor-bearing nude mouse model. It is effectively shown that the silent secretory clusterin reduces the hepatoma carcinoma nude mouse transplantation tumor growth speed. Through protein and gene analysis, it is effectively shown that the silent secretory clusterin enables the expression level of the silent secretory clusterin in a hepatoma carcinoma nude mouse transplantation tumor and mRNA of the silent secretory clusterin to be smaller than those of a blank control group and a negative control group.

Description

technical field [0001] The invention relates to the technical field related to Golgi secreted clusterin, in particular to a method for silencing secreted clusterin to inhibit liver cancer xenografting in nude mice. Background technique [0002] Liver cancer (Hepatocellular carcinoma, HCC) is one of the most common malignant tumors in the world, and its prevention and treatment are still difficult problems in the medical field. Early diagnosis of liver cancer is difficult and the prognosis is poor. Finding accurate and specific diagnostic and prognostic indicators will help in the prevention and treatment of HCC. The occurrence and development of HCC is a complex process of multi-cause, multi-gene, multi-step and multi-factor coordination, involving multi-signal pathways and gene regulation such as activation of oncogenes, inactivation of tumor suppressor genes, and reactivation of some oncogenes during the embryonic period. At present, surgical resection or liver transplant...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/85A01K67/027
CPCC12N15/85A01K67/0271A01K2207/12A01K2227/105A01K2267/0331
Inventor 姚登福钱琦郑文杰姚敏王理
Owner AFFILIATED HOSPITAL OF NANTONG UNIV
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