阅读说明：本技术 一种用于癌症预后检测试剂盒 (Kit for cancer prognosis detection ) 是由 叶定伟 于 2018-07-12 设计创作，主要内容包括：本发明公开了一种用于癌症预后检测试剂盒,试剂盒包括可检测ASPA、TMEM38B、PTGR2、TXLNB、ADGRE2表达水平的检测试剂；本发明提供了可高效预测癌症的预后的试剂盒。(The invention discloses a cancer prognosis detection kit, which comprises a detection reagent capable of detecting expression levels of ASPA, TMEM38B, PTGR2, TXLNB and ADGRE 2; the present invention provides a kit that can efficiently predict the prognosis of cancer.)

1. A detection kit for cancer prognosis, which comprises a detection reagent capable of detecting the expression level of ASPA, TMEM38B, PTGR2, TXLNB and ADGRE 2.

2. The kit of claim 1, wherein the detection reagent comprises a polynucleotide primer or probe.

3. The kit of claim 2, wherein the polynucleotide primer has a sequence as shown in SEQ ID NOs 1-10.

4. The kit of claim 1, wherein the cancer is selected from renal cancer.

5. The kit of claim 1, wherein said determination of the prognosis of cancer comprises the steps of:

a) detecting the mRNA expression level of ASPA, TMEM38B, PTGR2, TXLNB, ADGRE2 of the sample;

b) assessing the risk of cancer recurrence for the patient based on the expression level data obtained in step a).

6. The kit of claim 5, wherein in step b) the patient's risk of cancer recurrence is assessed by:

risk value ═ (1.6631 × ASPA expression level) + (0.2026 × TMEM38B expression level) + (0.1667 × PTGR2 expression level) + (-2.3969 × TXLNB expression level) + (-2.0618 × ADGRE2 expression level.

7. The use of a detection reagent for detecting the expression levels of ASPA, TMEM38B, PTGR2, TXLNB and ADGRE2 in the preparation of a product for cancer prognosis.

8. The use according to claim 7, wherein the cancer is renal cancer.

9. The use according to claim 7, wherein said determination of the prognosis of cancer comprises the steps of:

a) detecting the mRNA expression level of ASPA, TMEM38B, PTGR2, TXLNB, ADGRE2 of the sample;

b) assessing the risk of cancer recurrence for the patient based on the expression level data obtained in step a).

10. The use of claim 9, wherein in step b) the patient is assessed for risk of cancer recurrence by:

risk value ═ (1.6631 × ASPA expression level) + (0.2026 × TMEM38B expression level) + (0.1667 × PTGR2 expression level) + (-2.3969 × TXLNB expression level) + (-2.0618 × ADGRE2 expression level.

Technical Field

The invention relates to the field of medicine and health, in particular to a detection kit for cancer prognosis.

Background

Renal cancer is a malignant tumor that originates in the epithelial system of the urinary tubule of the renal parenchyma, and is known by the academic term as renal cell carcinoma. The kidney cancer accounts for about 2-3% of adult malignant tumor and 80-90% of adult malignant tumor. The incidence of diseases of countries or regions in the world is different, the incidence of diseases of developed countries is higher than that of developing countries in general, urban regions are higher than that of rural regions, more men are than women, the proportion of male patients to female patients is about 2: 1, the incidence of diseases can be found in all age groups, and the high incidence age is 50-70 years old. According to the statistics of the disease and death data of tumors in the trial-and-error cities and counties in China by the health statistical information center of the national cancer prevention and treatment research office and the Ministry of health, the disease rate of the kidney cancer in China is on the rising trend year by year, and the disease rate becomes the 10 th of the disease rate of the male malignant tumors in China by 2008.

Surgical treatment of early stage renal cancer is often the treatment of choice and is currently recognized as a cure for renal cancer. The kidney cancer patients in early stage can adopt nephron-sparing operation or radical nephrectomy. However, there is no recommendable adjuvant treatment regimen for patients with early and middle stage renal cancer after surgery to effectively prevent recurrence or metastasis. When patients develop relapse and metastasis, novel targeting regimens can be used for first or second line treatment of patients with metastatic renal cancer. However, this part of patients has a much poorer prognosis. In clinical work, how to distinguish high-risk renal cancer patients from low-risk renal cancer patients so as to purposefully adopt adjuvant therapy is still a great problem.

Functional genomics, also often referred to as post genomics, uses information and products provided by structural genomes to develop and apply new experimental approaches to switch biological research from single gene or protein research to multiple gene or protein simultaneous systematic research by comprehensively analyzing gene functions at the genomic or systematic level. This is a biological functional study of genome dynamics, which was carried out after the nucleotide sequence of the genome was clarified in a static state. The research content comprises gene function discovery, gene expression analysis and mutation detection. The functions of the gene include biological functions, such as phosphorylation modification of specific proteins as protein kinases; cytological functions, such as involvement in intercellular and intracellular signaling pathways; developmental functions, such as participation in morphogenesis. The adopted means comprises classical subtractive hybridization, differential screening, cDNA representation difference analysis, mRNA difference display and the like, but the technologies can not carry out comprehensive systematic analysis on genes, and new technologies come to the fore, including Systematic Analysis of Gene Expression (SAGE), cDNA microarray (cDNA microarray), DNA chip (DNAChip) and sequence tagged fragment display (suggested by the institute of China Zengbang, 20th ICG German Berlin) technology, microfluidic chip laboratory and the like.

A large number of genes related to kidney cancer prognosis have been found in the existing research, but most of the existing research only has a single data set, such as a paper in 2016 (Oncotarget), A four-gene signature prediction in clear-cell carcinoma, only one data set of TCGA, lack of verification and large bias. A Five-Gene signatures prediction characteristics in clinical science and journal of Computational and Mathematical methods in Medicine in clinical research Cell Carcinoma discloses that the expression of 5 genes in total, CKAP4, SLC40A1, OTOF, MAN2A2 and ISPD, is used as a model for predicting the Prognosis of Renal Clear Cell Carcinoma. The paper also discloses the use of this model to predict the outcome of renal cancer: the concentrated HR values were found to be 0.37, 95% CI: 0.30-0.46, HR value in test set 0.52, 95% CI: 0.43-0.62.

Although the prior art discloses a large number of kidney cancer-related genes, the use of kidney cancer-related genes for the evaluation of prognosis of kidney cancer in practical applications is limited. The prediction effect of the existing model still needs to be enhanced, and a model capable of predicting the risk of the kidney cancer more accurately needs to be established from a large number of kidney cancer genes.

Disclosure of Invention

In order to solve the technical problems, the invention provides a polygene kit.

In one aspect of the invention, a polygene detection kit for cancer prognosis is disclosed, and the kit comprises a detection reagent capable of detecting the expression levels of ASPA, TMEM38B, PTGR2, TXLNB and ADGRE 2.

Preferably, the detection reagent is a polynucleotide primer or probe.

Preferably, the sequence of the polynucleotide primer is shown in SEQ ID NO. 1-10.

Preferably, wherein the cancer is selected from renal cancer, preferably, the kit is for assessment of prognosis after renal cancer radical surgery.

Preferably, the cancer prognosis method of the present invention comprises the steps of:

(a) detecting the mRNA expression levels of ASPA, TMEM38B, PTGR2, TXLNB, ADGRE2 in the sample;

(b) assessing the patient's risk of cancer recurrence based on the expression level data obtained in step (a).

Preferably, in step (b) the patient is assessed for risk of cancer recurrence by:

(1.6631 × ASPA expression level) + (0.2026 × TMEM38B expression level) + (0.1667 × PTGR2 expression level) + (-2.3969 × TXLNB expression level) + (-2.0618 × ADGRE2 expression level), wherein said expression level is the mRNA expression value detected in step (a).

Preferably, the method for detecting the expression level of mRNA comprises: Affymetrix/Illumina chip detection, whole transcriptome shotgun sequencing, RT-PCR.

In another aspect of the invention, the use of the detection reagent for detecting the expression level of ASPA, TMEM38B, PTGR2, TXLNB and ADGRE2 in the preparation of a product for cancer prognosis.

Preferably, the present invention can be used for prognosis prediction of kidney cancer, and preferably, the present invention can be used for the evaluation of prognosis after radical treatment of kidney cancer.

Preferably, the cancer prognosis method comprises the steps of:

(a) detecting the mRNA expression levels of ASPA, TMEM38B, PTGR2, TXLNB, ADGRE2 in the sample;

(b) assessing the patient's risk of cancer recurrence based on the expression level data obtained in step (a).

Preferably, in step (b) the patient is assessed for risk of cancer recurrence by:

(1.6631 × ASPA expression level) + (0.2026 × TMEM38B expression level) + (0.1667 × PTGR2 expression level) + (-2.3969 × TXLNB expression level) + (-2.0618 × ADGRE2 expression level), wherein said expression level is the mRNA expression value detected in step (a).

Preferably, the method for detecting the expression level of mRNA comprises: Affymetrix/Illumina chip detection, whole transcriptome shotgun sequencing, RT-PCR.

Compared with the prior art, the technical scheme of the invention has the following advantages: the kit and the method can accurately predict the prognosis consequences of the cancer, and particularly have higher prediction value, important basis and clinical value and wide application prospect for high-risk diagnosis of the kidney cancer.

Drawings

In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and for those skilled in the art, other drawings can be obtained according to these drawings without creative efforts.

FIG. 1 is a 5-gene based gene decision model according to an embodiment of the present invention;

FIG. 2 is a ROC graph of 80 patients according to an embodiment of the present invention;

FIG. 3 is a graph of 80 patient survival plots for an embodiment of the present invention;

FIG. 4 is a graph of 515 patient ROC curves for an embodiment of the present invention;

FIG. 5 is a graph of 515 patient survival curves for an embodiment of the present invention.

Detailed Description

The conception, the specific structure and the technical effects of the present invention will be further described with reference to the accompanying drawings to fully understand the objects, the features and the effects of the present invention.

The terms "cancer" and "carcinoma" refer to or describe the physiological state in mammals that is typically characterized by abnormal or uncontrolled cell growth. Cancer and cancer pathology can be accompanied by, for example, metastasis, interference with the normal functioning of neighboring cells, release of cytokines or other secretory products at abnormal levels, suppression or aggravation of inflammatory or immune responses, neoplasia, precancerous lesions, malignancy, infiltration of surrounding or distant tissues or organs, such as lymph nodes, and the like. Particularly included are kidney cancers.

The term "prognosis" refers to the prediction of a medical outcome (medical output), such as poor or good outcome (e.g., likelihood of long-term survival); negative prognosis or adverse outcome includes the prediction of relapse, disease progression (e.g., tumor growth or metastasis or drug resistance), or death. Positive prognosis or good outcome includes prediction of disease improvement (e.g., disease-free state), improvement (e.g., tumor regression), or stabilization.

The ASPA of the invention is the same as the ASPA in SCI paper (Expression of aspartic enzyme (ASPA) and Anavan disease. journal book: Gene 2012 Sep 01; 505 (2)).

TMEM38B described in the present invention is similar to TMEM38B in SCI paper (Absence of the ER catalysis channel TMEM38B/TRIC-B dispersions Intracellular Calcium across and Dys regulated collagen Synthesis in Recessed Osteogenesis Imperfect, journal book PLoGenet.201607; 12(7))

The PTGR2 disclosed by the invention is the same as the PTGR2 in SCI paper (Targeting the 15-keto-PGE2-PTGR2a morphological evaluation system and subvalin experimental session journal book: Free Radic.biol.Med.2018 Feb 01; 115).

The TXLNB of the invention is TXLNB in the same SCI (RNA interference targeting CUG repeats in a mouse model of myonic dyrophy. journal volume: mol. the. 2013 Feb; 21(2))

The ADGRE2 of the present invention is similar to ADGRE2 in SCI paper (Membrane-association of EMR2/ADGRE 2-NTFis-defined by site-specific N-glycosylation, journal book: Sci Rep 2018Mar 14; 8 (1)).