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Establishing method of human pancreatic cancer gemcitabine-resistant cell line, and application of cell line

A technique for gemcitabine and method establishment, applied in the establishment of gemcitabine-resistant human pancreatic cancer cell line and the application field of the cell line, can solve problems such as unclear and complicated mechanism, achieve high drug resistance index, good stability, and efficient method Effect

Pending Publication Date: 2015-09-30
FUJIAN MEDICAL UNIV
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Problems solved by technology

The drug resistance of pancreatic cancer to GEM chemotherapy is the result of multi-factors and multi-factors, and the mechanism is complex and unclear

Method used

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  • Establishing method of human pancreatic cancer gemcitabine-resistant cell line, and application of cell line
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  • Establishing method of human pancreatic cancer gemcitabine-resistant cell line, and application of cell line

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Embodiment 1

[0012] Embodiment 1: a kind of method for establishing human pancreatic cancer resistant gemcitabine cell line, it comprises the following steps:

[0013] (1) Take 1×10 PANC-1 cell line in the exponential growth phase 8 Cells / L were inoculated in a 6-well plate, and GEM (Hengda Pharmaceutical Co., Ltd., dissolved in PBS buffer to a concentration of 100mmol / L, filtered and sterilized, aliquoted, and stored at -20°C) was added to a final concentration of 50nmol / L. The RPMI1640 culture medium (containing 15% FBS) of L, is placed 37 ℃, contains 5% CO2, cultivates in the incubator of saturated humidity;

[0014] (2) Observe the cell state. After 2-3 days, the cell state is poor, and many dead cells can be seen, so replace the same medium without GEM until the cell growth returns to normal (about 1 to 2 weeks); repeat the above steps , after repeated several times until the cell state is stable and there is no massive death, increase the final concentration of GEM by 25-50nmol / L; g...

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Abstract

The purpose of the invention is providing an establishing method of a human pancreatic cancer gemcitabine-resistant cell line, and an application of the cell line. The method comprises the following steps: putting a PANC-1 cell line in an exponential growth stage in a culture box, and culturing; observing the state of a cell until the growth of the cell goes back to normal; increasing the final concentration of GEM by 25-50nmol / L; gradually increasing the concentration until the final induction concentration is 1.9-2.1[mu]mol / L; and carrying out cloning culture on obtained drug-resistant cells through a limiting dilution process, selecting monoclonal cell growth holes with good cell state, and carrying out cloning culture. In the invention, the establishing method of the human pancreatic cancer gemcitabine-resistant cell line has the advantages of high efficiency and easy implementation; and the human pancreatic cancer gemcitabine-resistant cell line established in the invention has the advantages of high drug resistance index, good stability, and certain resistance to gemcitabine.

Description

technical field [0001] The invention relates to a method for establishing a gemcitabine-resistant cell line of human pancreatic cancer and the application of the cell line. Background technique [0002] Pancreatic cancer is a common malignant tumor of the digestive system. Due to its insidious onset, its surgical resection rate is low, its mortality rate is high, and its prognosis is poor. GEM has been used as the first-line drug for pancreatic cancer chemotherapy and has not been replaced for many years. However, due to the primary and secondary drug resistance of GEM, its efficacy is not satisfactory. The drug resistance of pancreatic cancer to GEM chemotherapy is the result of multi-factors and multi-factors, and the mechanism is complex and unclear. This study attempted to establish a GEM-resistant pancreatic cancer cell line and determine its biological characteristics. Contents of the invention [0003] The object of the present invention is a method for establishi...

Claims

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Application Information

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IPC IPC(8): C12N5/09C12Q1/02C12R1/91
Inventor 谢捷明黄鹤光王丛菲杨爱琴付明娟
Owner FUJIAN MEDICAL UNIV
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