Unlock instant, AI-driven research and patent intelligence for your innovation.

Application of pak5 in cancer diagnosis, prognosis, treatment and drug screening

A cancer and prognosis technology, applied in the field of genetic diagnosis, can solve the problems of early diagnosis and prognosis follow-up biomarkers that have not yet been found

Active Publication Date: 2020-09-25
XUZHOU MEDICAL UNIV
View PDF3 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, early diagnosis of kidney cancer patients is very important for distinguishing pathological types of the disease, indicating the degree of malignancy, monitoring chemotherapy resistance, and judging prognosis and recurrence. However, there is no suitable early diagnosis for kidney cancer or even most solid tumors. and prognostic follow-up biomarkers
However, existing studies have shown that certain proteins may become a potential new approach for early diagnosis and treatment of kidney cancer

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Application of pak5 in cancer diagnosis, prognosis, treatment and drug screening
  • Application of pak5 in cancer diagnosis, prognosis, treatment and drug screening
  • Application of pak5 in cancer diagnosis, prognosis, treatment and drug screening

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] Differential expression of genes of the present invention

[0053] 1. Sample collection:

[0054] Kidney cancer tissue samples: We collected kidney cancer patients, all of whom underwent surgical treatment, including 34 pairs of kidney cancer and their adjacent normal kidney tissues. All patients were diagnosed with renal cell carcinoma by pathological examination. Other enrollment conditions are: all patients have not received any treatment before admission; no other malignant tumors; no other hormone-related diseases; complete clinical data.

[0055] 2. Validation at the mRNA level

[0056] 2.1 Extract tissue RNA:

[0057] Total tissue RNA was extracted using Trizol one-step method, and the concentration of RNA solution was determined by reading the absorbance value (OD value) at 260nm and 280nm through Nanodrop ND-1000. 1% formaldehyde denatured agarose gel electrophoresis, observed under ultraviolet transmission light, check the integrity of the RNA.

[0058]...

Embodiment 2

[0071] Differential expression of PAK5 protein

[0072] 1. Sample collection:

[0073] Kidney cancer tissue samples: We collected kidney cancer patients, all of whom were surgically treated, including 307 kidney cancer specimens and 34 tissue microarrays of normal kidney tissue. All patients were diagnosed with renal cell carcinoma by pathological examination. Other enrollment conditions are: all patients have not received any treatment before admission; no other malignant tumors; no other hormone-related diseases; complete clinical data.

[0074] 2. Detect the expression level of PAK5 according to the following method:

[0075] (1) Blocking peroxidase: take the tissue chip for dewaxing, dewaxing until hydration (the dewaxing process is as follows: xylene Ⅰ 10 min → xylene Ⅱ 10 min → absolute ethanol 5 min → 95% ethanol 5 min → 85% ethanol 5 min → 75% ethanol 5min), with 3% H 2 o 2 The solution blocked endogenous peroxidase in the dark and incubated at room temperature ...

Embodiment 3

[0092] Inhibition of PAK5 gene expression

[0093] 1. Design and synthesis of siRNA

[0094] siRNA sequence against PAK5: 5'-CAAAGTCTTCGTACCTGAATC-3'

[0095] Negative control sequence: 5'-UCUACUCUUUCUAGGAGGUUGUGA-3'

[0096] The above siRNA sequences and the negative control siRNA sequence (siRNA-NC) were provided by Shanghai Aibosi Biotechnology Co., Ltd.

[0097] 2. Culture and transfection of renal cancer cells

[0098] 2.1 Cell culture

[0099] 786-O cells were cultured in RPMI 1640 medium (containing 10% fetal bovine serum, 100U / ml penicillin, 100g / ml streptomycin), ACHN cells were cultured in DMEM medium (containing 10% fetal bovine serum, 100U / ml penicillin, 100g / ml streptomycin), all at 37°C, 5% CO 2 Continuous cultivation in a humidified incubator.

[0100] 2.2 Cell transfection

[0101] (1) 24 hours before transfection, inoculate 1-2*10 5 cells, the cell confluency was 30-50% at the time of transfection. When plating, the cells should be digested and mixe...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention discloses applications of PAK5 to diagnosis, prognosis and treatment of cancer as well as drug screening, and comprises applications of PAK5 gene or PAK 5 protein to preparation of a reagent or a kit to diagnosis or prognosis of cancer. A method for screening candidate compounds for treating cancer is as follows: if expression level and / or activity of PAK5 of cells in a test group is higher than the expression level and / or activity of PAK5 of cells in a control group, the tested compound is a candidate compound for treating cancer with promotion effects for expression and / or activity of PAK5. A molecular marker for diagnosis of kidney cancer is discovered, the molecular marker can be used for determination of generation of kidney cancer in an early stage, and survival rate of patients is provided. In addition, by means of prediction of prognosis of patients, meaningful information is provided for patients to determine treatment schemes and strategies. A therapeutic which comprises an inhibitor of the PAK5 gene or protein can be used as a new therapeutic for kidney cancer.

Description

technical field [0001] The invention belongs to the field of molecular biology, especially gene diagnosis, especially the application of PAK5 in cancer diagnosis, prognosis, treatment and drug screening. Background technique [0002] Renal cell carcinoma (RCC), also known as kidney cancer, accounts for about 3% of all malignant tumors (Jemal, A., et al., Cancer statistics, 2010. CA Cancer J Clin, 2010.60 (5): p .277-300), which originates from the renal tubular epithelium, and is mainly composed of clear cell carcinoma, chromophobe carcinoma, and papillary carcinoma, among which clear cell carcinoma of the kidney is the most common, accounting for about 4 / 5 of the total (Zeng , Z., et al., A study exploring critical pathways in clear cell renal cell carcinoma. Exp Ther Med, 2014.7(1): p.121-130.). In recent years, the incidence of renal cancer has been increasing year by year, and the age is gradually decreasing. According to statistics, the incidence is increasing at an an...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6886G01N33/574G01N33/68C12Q1/02
CPCC12Q1/6886C12Q2600/118C12Q2600/136C12Q2600/158G01N33/5011G01N33/57438G01N33/68G01N2800/52
Inventor 裴冬生潘垚捷刘旭魏璐璐霍福春任鑫刘慧
Owner XUZHOU MEDICAL UNIV