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Method for detecting influence of targeted drug with multiple target points on melanoma cells

A melanoma cell and detection method technology, which is applied in the field of detection of the impact of targeted drugs on the survival and metastasis of melanoma cells, and can solve the problems that single-target inhibitors cannot meet the needs of tumor treatment and melanoma curative effect is weak

Inactive Publication Date: 2015-11-25
HUZHOU CENT HOSPITAL
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Problems solved by technology

However, Eisen et al. [18] conducted a phase II clinical trial on 37 patients with unresectable melanoma and distant metastases and found that sorafenib monotherapy had little effect on melanoma.
[0006] At present, targeted therapy has become one of the hottest areas of tumor treatment research, and the occurrence and development of tumors is the result of the participation of multiple genes and kinases. Therefore, single-target inhibitors are far from being sufficient for tumor treatment.

Method used

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  • Method for detecting influence of targeted drug with multiple target points on melanoma cells
  • Method for detecting influence of targeted drug with multiple target points on melanoma cells

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Embodiment Construction

[0016] Below, the technical solution of the present invention will be described in detail through specific examples.

[0017] 1. Effects of drug treatment on the proliferation inhibition rate, cell cycle and apoptosis rate of melanoma cell A375 cells

[0018] ①Different tumor cells in logarithmic growth phase were digested with 0.25% trypsin to make single cell suspension.

[0019] ② Adjust the cell concentration (1000-10000 cells per well, determined according to the actual growth of different cells) and inoculate in a 96-well culture plate, 0.2ml per well.

[0020] ③Cultivate at 37°C and 5% CO2 for 24 hours. After the growth reaches 70-80%, remove the original culture solution, prepare the treatment drug with DMSO (dimethyl sulfoxide) to form a mother solution, and then stir with the corresponding cell culture solution Diluted to 5 μmol / L, 10 μmol / L, 20 μmol / L, 40 μmol / L, respectively added to 96-well plate, set blank control group, cell control group, sorafenib group, conv...

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Abstract

The invention discloses a method for detecting influence of a targeted drug with multiple target points on survival and metastasis capacities of melanoma cells. The detection method comprises the following steps: treatment of melanoma cells; RT-PCR detection of expression of a cell metastasis-associated gene PERK; western blot detection of expression of a cell metastasis-associated protein PERK; observation of invasion and metastasis capacity of each group of melanoma cells through Transwell experiments; and integration of data and analysis of effects of the targeted drug with multiple target points on survival and metastasis capacities of melanoma cells. The detection method can systematically detect influence of the targeted drug on survival and metastasis capacities of melanoma cells.

Description

technical field [0001] The invention relates to the technical field of medical detection methods, in particular to a detection method for detecting the influence of multi-target targeted drugs on the viability and metastasis of melanoma cells. Background technique [0002] Melanoma is a malignant tumor originating from melanocytes in the skin, mucous membranes, eyes, and pigmented areas of the central nervous system. It is the skin cancer with the highest mortality rate. It is estimated that in 2009, there were 68,720 melanoma cases in the United States, accounting for 92.1% of skin cancer cases, and 8,650 deaths, accounting for 74.6% of skin cancer deaths, and the annual prevalence rate is rising rapidly. The cause of melanoma is still unclear, and the treatment options are mainly surgery and radiation therapy, supplemented by drug therapy, but the cure rate for middle and advanced patients is very low. The reason is that advanced tumors are prone to metastasis. Tumor met...

Claims

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Application Information

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IPC IPC(8): G01N33/15
Inventor 陆群英吴原戴利成
Owner HUZHOU CENT HOSPITAL
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