A method for inducing transformation of adult liver stem cells into highly metastatic liver cancer cells and corresponding cells

A technology for liver stem cells and liver cancer cells, which can be used in tumor/cancer cells, chemical instruments and methods, non-embryonic pluripotent stem cells, etc. It can solve the problems of uncertain tumor formation time, unstable liver cancer cells, and low success rate.

Active Publication Date: 2021-02-19
SUN YAT SEN MEMORIAL HOSPITAL SUN YAT SEN UNIV
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Problems solved by technology

It is not yet clear whether malignantly transformed normal cells acquire tumorigenicity and the ability to metastasize
At the same time, the current model for studying lung metastasis of liver cancer is limited. Human-derived liver cancer is poor in lung metastasis, and animal orthotopic liver transplantation is limited to immunodeficient mice, with low success rate and high mortality.
[0006] In the prior art, rat adult liver stem cells that have not been transfected with any gene can spontaneously form tumors after 8-12 cycles of continuous subculture, but the treatment time is long, the spontaneous tumor formation rate is low, and malignant transformation cells without lung metastasizing ability
[0007] Using Notch1 gene to transfect adult liver stem cells or hepatocytes, without serial passage, can only form tumors, and the tumor formation time in the liver of nude mice is long, with low tumorigenicity, and no lung metastasis has been reported
[0008] If other genes are used to transfect adult liver stem cells or hepatocytes without continuous passage, although tumors can be formed, the time of tumor formation is uncertain, the tumorigenicity is low, and no lung metastasis is seen.
[0009] The disadvantage of the above method is that the obtained liver cancer cells are unstable, and the tumor formation time is long, the tumorigenicity is poor, and they do not have the ability to metastasize to the lungs.

Method used

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  • A method for inducing transformation of adult liver stem cells into highly metastatic liver cancer cells and corresponding cells
  • A method for inducing transformation of adult liver stem cells into highly metastatic liver cancer cells and corresponding cells
  • A method for inducing transformation of adult liver stem cells into highly metastatic liver cancer cells and corresponding cells

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Embodiment 1

[0038] This example provides a method for inducing transformation of adult liver stem cells into highly metastatic liver cancer cells, and provides a stable liver cancer cell with short tumor formation time, good tumorigenicity, and lung metastatic ability.

[0039] The concrete experimental scheme of this embodiment is as follows:

[0040] S1. Expansion and culture of Fisher 344 rat adult liver stem cells WB-F344

[0041](1) Recovery of WB-F344: routinely sterilize the ultra-clean workbench, add 10mL DMEM medium into a sterile 15mL centrifuge tube, take out a tube of frozen rat adult hepatic stem cells WB-F344 from the liquid nitrogen tank, and immediately Place in a water bath at 37°C and oscillate to observe the thawing of the cell suspension. After about 2 / 3 of the thawing, spray the cryopreservation tube with 75% alcohol and put it into an ultra-clean bench. Use a straw to remove the cell suspension. Transfer to a centrifuge tube containing 10mL of medium, blow it evenly...

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Abstract

The present invention provides a method for inducing the transformation of adult liver stem cells into highly metastatic liver cancer cells and the stable liver cancer cells screened by the method. The method includes the following steps: S1: amplifying, culturing and preserving rat adult liver stem cells; S2: Using gene transfection technology to transfer the lentiviral vector containing Notch1 gene into adult rat hepatic stem cells to obtain rat adult hepatic stem cells stably overexpressing Notch1; S3: Using rat adult hepatic stem cells stably overexpressing Notch1, using Selective conditional culture method was used to cultivate them to further differentiate in vitro, and to screen out hepatocellular carcinoma cells with strong tumorigenicity and stability; Hepatocarcinoma was formed in the liver, accompanied by the spontaneous formation of bilateral lung metastases, resulting in the acquisition of highly metastatic hepatocellular carcinoma cells. The stable liver cancer cells screened by the method of the invention have short tumor forming time, good tumorigenicity and lung metastatic ability.

Description

technical field [0001] The invention relates to a method for inducing transformation of adult liver stem cells into highly metastatic liver cancer cells and corresponding cells. Background technique [0002] Liver cancer is a common malignant tumor in my country. It has the characteristics of insidious onset, rapid progression, easy early hematologic metastasis, difficult to cure, and poor prognosis. Although great progress has been made in the early diagnosis, surgical resection and comprehensive treatment of liver cancer, the five-year survival rate of liver cancer has not improved significantly in the past ten years. Postoperative recurrence of liver cancer has always been the main reason restricting the efficacy of liver cancer treatment. In recent years, it has been found that tumor stem cells are closely related to the formation, development, metastasis, recurrence and drug resistance of tumors. The theory of cancer stem cells believes that most tumors originate from...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/10
CPCC07K14/47C12N5/0693C12N2506/03C12N2510/00
Inventor 刘超施祥德吴文睿许磊波余先焕张锐
Owner SUN YAT SEN MEMORIAL HOSPITAL SUN YAT SEN UNIV
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