Method for inducing adult liver stem cells for transforming to high metastatic liver cancer cells and corresponding cells

Abstract

The invention provides a method for inducing adult liver stem cells for transforming to high metastatic liver cancer cells and the stable liver cancer cells screened by the method. The method comprises the following steps: S1: performing amplification, culture and preservation of mice adult liver stem cells; S2: employing a gene transfection technology for transferring a lentiviral vector containing the Notch1 gene into the mice adult liver stem cells to obtain the mice adult liver stem cells for stable over-expression of the Notch1; S3: using the mice adult liver stem cells for stable over-expression of Notch1, employing a selective condition culture method for culture, performing in-vitro further differentiation, screening the liver cancer cells having strong and stable tumorigenic capability; and S4: employing the screened liver cancer cells, through an in-situ liver transplantation tumor model having same species, liver cancer is formed on liver, double-lung metastasis tumor is spontaneously formed, so that the high metastatic liver cancer cells can be obtained. The screened stable liver cancer cells by the method of the invention have the advantages of short tumor formation time and good tumorigenicity, and have lung metastasis capability.

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...

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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.

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