Polygene detection kit for cancer prognosis

A detection kit and detection reagent technology, applied in the field of medicine and health, can solve the problems that the prediction effect needs to be enhanced, the evaluation of prostate cancer is limited, etc., and achieve the effect of high prediction value

Active Publication Date: 2019-11-19
FUDAN UNIV SHANGHAI CANCER CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Although a large number of prostate cancer-related genes have been disclosed in the prior art, the use of prostate cancer-related genes in evaluating prostate cancer in practical applications is limited
The p...

Method used

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  • Polygene detection kit for cancer prognosis
  • Polygene detection kit for cancer prognosis
  • Polygene detection kit for cancer prognosis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Example 1 Establishment of Prostate Cancer Recurrence and / or Survival Consequence or Prognosis Assessment Model

[0036] By searching and screening 149 prostate cancer patients from the Gene Expression Omnibus (GEO) database, the corresponding microarray data were extracted. According to the research progress of prostate adenocarcinoma, 522 potential target genes were identified, and the Lasso Cox regression model was used to analyze the mRNAs closely related to the biochemical recurrence of these patients. Finally, a gene decision model based on 15 mRNAs was determined and constructed (see figure 1 ). And according to the risk of the decision model, the following scoring formula was used to obtain the risk value, and the patients were divided into high-risk group and low-risk group for biochemical recurrence after prostate cancer surgery.

[0037] Risk value=(0.4237×MCM2 expression level)+(0.1915×GATM expression level)+(-0.1810×PTGDS expression level)+(0.2629×ETV1 ex...

Embodiment 2

[0040] Example 2 Multigene Kit for Prostate Cancer Prognosis

[0041] The multigene kit used for the prognosis of prostate cancer in this embodiment includes:

[0042] Total RNA extraction reagent Trizol;

[0043] Chloroform (chloroform);

[0044] isoamyl alcohol;

[0045] Anhydrous ethanol;

[0046] DEPC water (DD1005);

[0047] Ethylene glycol pyrophosphate (DEPC);

[0048] Anti-RNase solution (RNaseZap);

[0049] Reverse transcription kit;

[0050] iQ SYBR Green Supermix;

[0051] The polynucleotide primer whose sequence is SEQ ID NO: 1-30.

[0052] After using the kit to perform real-time quantitative PCR (qRT-PCR) detection of each gene, the initial data result is represented by the Ct value (cycle threshold), that is, the number of cycles required for the fluorescent signal in each reaction system to reach the set threshold. The Ct value of each sample has a linear relationship with the logarithm of the initial copy number of the sample, and the higher the initi...

Embodiment 3

[0055] Example 3 Preparation and pretreatment of sample RNA

[0056] (1) For the tissue RNA extraction experiment, soak the tip and tweezers in DEPC water overnight, then sterilize under high temperature and high pressure, pre-cool the centrifuge at 4°C, and wipe the test bench, pipette gun, gloves, etc. with anti-RNase Zap.

[0057] (2) Tissue homogenate Take 50-100mg tissue sample, mince it slightly with a sterile scalpel blade, put it into a 1.5mL EP tube, add 500μL Trizol reagent, fully homogenize with an electric tissue grinder, and then add 500μL Trizol reagent .

[0058] (3) Add 0.2mL chloroform to every 1mL Trizol reagent homogenate sample, and tightly cap the EP tube. Shake vigorously for 15 seconds, and place at room temperature for 3 minutes. Centrifuge at 12,000 rpm at 4°C for 15 minutes (pre-cool the centrifuge). After centrifugation, the mixed system will be divided into a colorless aqueous phase in the upper layer, a protein in the middle layer and a red phen...

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Abstract

The invention discloses a polygene detection kit for cancer prognosis. The kit comprises a detection reagent capable of detecting the expression levels of MCM2, GATM, PTGDS, ETV1, CASP3, NOTCH3, WNT5A, GLIS1, TNFRSF10B, COL4A6, MMP19, MSR1, PALLD, CCNG2 and TMPRSS2. The present invention provides a kit capable of efficiently predicting prognosis of cancer.

Description

technical field [0001] The invention relates to the field of medical and health fields, in particular to a multigene detection kit for cancer prognosis. Background technique [0002] Radical prostatectomy is one of the important methods for the treatment of localized prostate cancer. However, after radical prostatectomy, biochemical recurrence still occurs in quite a few patients. When a patient has biochemical recurrence, it is necessary to clarify whether clinical recurrence has occurred, and whether it is local recurrence, regional lymph node metastasis or distant metastasis. In clinical work, the assessment of biochemical recurrence after radical prostatectomy is often a headache, which leads to the lack of precision in its treatment. [0003] Functional genomics (Functional genomics) is also often referred to as postgenomics (Postgenomics), which uses the information and products provided by the structural genome to develop and apply new experimental methods, by com...

Claims

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Application Information

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IPC IPC(8): C12Q1/6886
CPCC12Q1/6886C12Q2600/118C12Q2600/158
Inventor 邵宁
Owner FUDAN UNIV SHANGHAI CANCER CENT
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