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Biomarker in diagnosis of liver cancer and kit containing biomarker in diagnosis of liver cancer

A biomarker and diagnostic kit technology, applied in the field of RPS3 detection, can solve the problem of unclear molecular mechanism of liver cancer and other problems

Active Publication Date: 2020-05-15
PEKING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The evolution of normal liver cells into liver cancer cells needs to go through multiple pathological stages, involving a variety of molecular events, but the detailed molecular mechanism of liver cancer development is still unclear

Method used

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  • Biomarker in diagnosis of liver cancer and kit containing biomarker in diagnosis of liver cancer
  • Biomarker in diagnosis of liver cancer and kit containing biomarker in diagnosis of liver cancer
  • Biomarker in diagnosis of liver cancer and kit containing biomarker in diagnosis of liver cancer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0189] Example 1 Effect of RPS3 overexpression or knockdown on the function of liver cancer cells

[0190] HepG2 and SMMC-7721 liver cancer cell lines were used to knock down RPS3 at the cell line level to verify the role of RPS3 in the proliferation, migration, invasion, EMT and clone formation ability of liver cancer cells.

[0191] Result description:

[0192] We transferred the adenovirus sh-RNA of RPS3 into HepG2 cells, and the knockdown effect was verified by Q-PCR and Western blot, which proved that the knockdown of RPS3 expression was achieved in the corresponding cells.

[0193] Normal HepG2 cells were used as the control group, and RPS3-knockdown HepG2 cells were used as the test group. After the two groups of cells were cultured, cell proliferation, clone formation, cell cycle, wound healing, and small hole mobility were detected.

[0194] For the results of cell proliferation ability and colony formation, see Figure 4 and Figure 5 . From Figure 4 It can be ...

Embodiment 2

[0200] Effect of Knocking Down RPS3 on Tumor Growth in Example 2 Nude Mouse Model

[0201] Six BALB / c mice (female, 6-8 weeks old) were used in each experiment, and RPS3-knockdown SMMC-7721 cells and normal SMMC-7721 cells were injected subcutaneously to observe the effect of RPS3 on tumorigenesis at the animal level. The impact of sex, and immunohistochemical analysis of tumor tissue to detect the expression of RPS3.

[0202] Collect RPS3 knockdown virus stable strain cells, with 3 × 10 6 The amount of cells / mouse / day was injected subcutaneously for 2 weeks, and the size of the tumor was observed and measured with a caliper. After 4 weeks, the mice were sacrificed, and the tumors were removed for immunohistochemical analysis.

[0203] After the tumor tissues of nude mice were dewaxed and dehydrated at 62°C for 2 hours, antigen retrieval was performed at 97°C for 20 minutes in citrate buffer, 3% H 2 o 2 Block endogenous peroxidase at room temperature for 10 minutes, incuba...

Embodiment 3

[0206] Example 3 Effect of FMHM on the expression of RPS3 and the function of liver cancer cells

[0207] The SMMC-7721 cells and HepG2 cells were divided into 1×10 4 The cells / well were plated, and after the cells adhered to the wall, gradient drug treatment was performed. A stock solution of FMHM was prepared in DMSO, followed by stepwise dilution with simple medium. The drug concentration gradient of FMHM was 1000 μM, 250 μM, 62.5 μM, 15.625 μM, 3.906 μM, 0.977 μM, 0.244 μM, 0.061 μM, 0.0152 μM, 0.0038 μM, 0.000954 μM. After 48 hours, the cell growth curve was drawn. Calculate the IC50 (half maximal inhibitory concentration, half inhibitory concentration) of FMHM according to the concentration and cell survival rate, and obtain the optimal concentration of FMHM for different cell lines: SMMC-7721 cells are 32.42 μM, HepG2 cells are 15.8 μM, and this concentration is used as Subsequent experimental doses.

[0208] Different liver cancer cells were treated with different ...

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Abstract

The invention provides a biomarker in diagnosis of liver cancer and application of the biomarker in diagnosis of liver cancer in production of a diagnostic reagent of liver cancer. It is found throughresearch that overexpression of RPS3 can promote liver cancer cell proliferation, clone formation, migration, invasion and the transformation capability of epithelial cells into mesenchymal cells, and on the contrary, knock-down of RPS3 can inhibit liver cancer cell proliferation, clone formation, migration, invasion and the transformation capability of epithelial cells into mesenchymal cells; after a liver cancer cell strain with knock-down RPS3 is subcutaneously injected into nude mice, the tumorigenicity is reduced; it is also found in people that compared with expression of RPS3 in para-carcinoma tissue, expression of RPS3 in liver cancer tissue is high, and high expression of the RPS3 affects the prognosis survival rate of patients to a certain extent; and on the basis, the biomarkerin diagnosis of liver cancer and application of the biomarker in diagnosis of liver cancer in production of the diagnostic reagent of liver cancer are provided. The biomarker is a RPS3 gene, mRNA ofthe RPS3 gene, or protein or a protein fragment encoded by the RPS3 gene, and the biomarker can be used as a medicine target for screening liver cancer medicines.

Description

technical field [0001] The present invention relates to the field of biomedicine, in particular to biomedical technology for the diagnosis and treatment of liver cancer, in particular to the application of the RPS3 gene and its encoded protein as a diagnostic biomarker and therapeutic target for liver cancer, and RPS3 as the detection object. Application of liver cancer diagnosis kit and compound 4-hydroxy-2-methylene butyrate (5-formylfuran-2-methyl) ester in preparation of medicine for treating liver cancer. Background technique [0002] Primary liver cancer (PLC) is a primary liver disease with a high mortality rate and one of the most common malignant tumors worldwide. According to incomplete statistics, the annual increase of liver cancer in the world exceeds 600,000, and 55% of new liver cancers occur in China. Primary liver cancer can be divided into massive liver cancer, nodular liver cancer, and diffuse liver cancer in terms of pathology; hepatocellular carcinoma (...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886G01N33/68G01N33/574
CPCC12Q1/6886C12Q2600/118C12Q2600/158G01N33/57438G01N33/68
Inventor 韩丽敏赵丽君胡克新童坦君陈军
Owner PEKING UNIV
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