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Rat liver cancer cell line with high metastatic potential, preparation method and application thereof

A high-metastasis technology of rat liver cancer cells, applied in the field of rat liver cancer cell lines and its preparation, can solve the problems of liver cancer affecting the curative effect of surgery

Inactive Publication Date: 2018-03-06
HEBEI MEDICAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the postoperative recurrence and metastasis of liver cancer directly affect the curative effect of surgery, and the metastasis and recurrence of liver cancer have become the main obstacle to further prolong the survival and improve the prognosis of liver cancer patients.

Method used

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  • Rat liver cancer cell line with high metastatic potential, preparation method and application thereof
  • Rat liver cancer cell line with high metastatic potential, preparation method and application thereof
  • Rat liver cancer cell line with high metastatic potential, preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0059] Example 1: Preparation of liver cancer cell line Wrh-f2 with high metastatic potential

[0060] The male Wistar rats used in this example were provided by Beijing Weitong Lihua Experimental Animal Technology Co., Ltd.

[0061] The DEN (diethylnitrosamine) used in this example, the Chinese name is N-diethylnitrosamine, was purchased from Sigma Company; the NMOR (N nitrosomorpholine), the Chinese name was N-nitrosomorpholine, was purchased from Sigma Company.

[0062] (1) Preparation of liver cancer cells

[0063] 1. Select 40 male Wistar rats with a body weight of 130~150g, and inject DEN (prepared with normal saline, concentration 10mg / mL) intraperitoneally at 100mg / kg on the first day, and give drinking water containing 100ppm NMOR starting from the second day (prepared with sterilized tap water), let them drink freely, use them for 5 consecutive days a week, and then switch to sterile water for feeding on the next 2 days, and then supply NMOR drinking water and steri...

Embodiment 2

[0075] Example 2: Analysis of biological characteristics of high metastatic potential cell line Wrh-f2 and comparison with low metastatic potential cell line Wrh-s2:

[0076] 1) Observation of cell morphology:

[0077] The two cell lines were observed with an inverted phase-contrast microscope, and the results are as follows:

[0078] ① Under the microscope, 97% of the cells were polygonal cells, 3% were round cells, and no fibroblasts were found, indicating that the cell morphology was relatively uniform, which was in line with the characteristics of liver cancer cells;

[0079] ②. The cultured cells showed large nuclei, multinucleated cells, abnormal mitotic phases, and the characteristics were the same as those of the patient's liver cancer tissue (see Figure 4 );

[0080] ③. When the density is low, the cells grow in a clone-like manner. When the density is high, the cells grow overlappingly. The naked eye can see a small tissue block-like structure, and the cell bounda...

Embodiment 3

[0120] Example 3 Establishment of Transplanted Liver Cancer Model

[0121] 120-130g 20 Wistar rats, male, made cell paste from 2 strains of cells, inoculated 0.04ml under the liver capsule of each rat, and observed the growth and metastasis of liver tumors.

[0122] 1) Two weeks later, liver tumor growth was seen in both groups, and lung metastasis occurred in 70% of rats inoculated with Wrh-f2 cell line, and in 10% of rats inoculated with Wrh-s2 cell line.

[0123] 2) Hepatic HE staining shows polygonal cancer cells with rich cytoplasm, round or oval nuclei, obvious nucleoli, obvious atypia, and frequent mitotic phases; occasionally giant tumor cells are seen, and the cells are arranged in cords and strips with fibers Separation, rich interstitial blood vessels, consistent with the morphology of human liver cancer.

[0124] 3) HE staining of lung cells in Wrh-f2 group showed obvious infiltration of liver cancer cells.

[0125] The establishment experiment of the transplante...

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Abstract

The invention discloses a rat liver cancer cell line with high metastatic potential. The cell line is named rat liver cancer cell Wrh-f2, and was preserved in the China Center for Type Culture Collection on October 21, 2015, and the preservation number is CCTCC C2015174. The invention also provides the preparation method of the cell line and its application in the preparation of transplanted liver cancer models. In order to obtain cells with a high transfer rate, the present invention adopts an improved rat liver cancer induction method, and then takes the well-grown tumor tissue and adopts a direct cell monolayer culture method for primary culture to obtain rat liver cancer cells, and then uses Transwell Cells with high metastatic potential are screened out by the small chamber and used to prepare a transplanted rat liver cancer model, which will be very beneficial to analyze the mechanism of liver tumor metastasis. In addition, we cultured and purified cells by cloning, selected well-growing cells with high metastatic potential and preserved them, and through a series of experiments, we proved that the screened Wrh‑f2 cell line is a liver cancer cell line with good tumorigenic properties.

Description

technical field [0001] The invention relates to a biological cancer cell line and its preparation technology, in particular to a rat liver cancer cell line with high transfer potential, its preparation method and application. Background technique [0002] With the continuous development of liver cancer-related research, as the basis of preclinical in vivo research, the establishment of liver cancer animal models is also making progress. The induced liver cancer model is still the most widely used animal model in liver cancer research because it can better imitate the process of human liver cancer. However, the animal model of induced liver cancer has the disadvantages of long period (2-5 months), high cost, low success rate, and difficulty in obtaining a model with relatively uniform disease course and tumor size at the same time, and its application has been limited. The liver cancer model made by transplantation has the advantages of simple animal breeding, simple transpl...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/09
Inventor 张焕铃刘树锋宋蕾王昭
Owner HEBEI MEDICAL UNIVERSITY
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