Urine-based method and kit for diagnosing relapse risk of bladder cancer patient

A bladder cancer and risk technology, applied in the field of biomedical detection, can solve the problems of high operator's technical dependence, low detection rate of urine exfoliated cells morphological examination, and high price.

Inactive Publication Date: 2012-04-25
SHANGHAI INST OF ONCOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

B-ultrasound is easily affected by other diseases of the bladder, such as bladder polyps, bladder submucosal hemorrhage, etc., and it is easy to miss tumors <5mm or cases that are spread on the mucosal surface and do not protrude into the cavity; while cystoscopy is an invasive examination, At the same time, it is highly dependent on operator skills, and the price is more expensive; in addition, the detection rate of urine exfoliated cell morphology is low, and the detection rate of early stage Ta and T1 bladder cancer is 0

Method used

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  • Urine-based method and kit for diagnosing relapse risk of bladder cancer patient
  • Urine-based method and kit for diagnosing relapse risk of bladder cancer patient
  • Urine-based method and kit for diagnosing relapse risk of bladder cancer patient

Examples

Experimental program
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Effect test

Embodiment 1

[0075] Example 1. Establishment of high-throughput methylation profiles of bladder cancer cell lines and normal bladder mucosa at the genome level

[0076] The present invention conducts high-throughput genomic analysis on two cases of normal human bladder mucosa (obtained from Urology Department of Zhongshan Hospital) and two cases of bladder cancer cell lines (5637 (ATCC: HTB-9), T24 (ATCC-4)). Establishment of methylation profiles.

[0077] Step 1: Extract the DNA of the above-mentioned bladder cancer cells and normal tissues, and use the MBD (methylation binding protein domain) affinity chromatography column to enrich the high, medium and low methylated DNA fragments step by step; Step 2: To collect The highly methylated DNA fragments were added with SOLEXA adapters and sent to Shanghai Bohao Biotechnology Co., Ltd. for SOLEXA high-throughput sequencing. The bladder cancer tumor cell group and the normal bladder tissue group obtained 3 million reads respectively. After bi...

Embodiment 2

[0078] Example 2. Determination of the methylation profile of bladder cancer-related genes

[0079] Step 1: Preliminary screening of large gene groups in small-scale clinical samples

[0080] Further screened from the above 1627 highly methylated regions associated with gene promoters in tumor cell lines to obtain the promoter-related regions of the top 104 genes with the largest difference in methylation value (sorted according to P value, select the region related to the normal bladder The top 104 genes with the highest degree of methylation compared to tissues).

[0081] The regions related to the promoters of the top 104 genes with the largest methylation differences were initially screened for methylation differences between bladder cancer cell lines and normal population urine samples (urine sediment).

[0082] Genomic DNA extraction methods are commonly used protease cleavage and phenol / chloroform extraction. Bisulphite (bisulphite) treatment is as follows: 1ug genomi...

Embodiment 3

[0113] Embodiment 3, clinical and follow-up analysis and information statistics

[0114] The clinical urine samples of 212 bladder cancer patients collected earlier were taken from untreated bladder cancer patients diagnosed by cystoscopy and pathological examination; 157 of them were primary cases and 55 were recurrent cases, accounting for 25.94%; these patients have been treated with surgery (total cystectomy, partial cystectomy, electroablation of bladder tumor, combined with chemotherapy or radiotherapy), and no local residual tumor lesions in the bladder have been found by cystoscopy. After excluding 12 uncontactable cases from 157 primary cases, and following up the remaining 145 cases, 37 patients relapsed, accounting for 25.5%.

[0115] The correlation between gender, clinical stage, case grade, muscle invasion, primary recurrence and methylation of LMX1A and VAX1 in 212 cases of bladder cancer was analyzed by univariate and multivariate logistic regression. The resu...

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Abstract

The invention relates to a urine-based method and kit for diagnosing the relapse risk of a bladder cancer patient. The invention discloses two methylation-sensitive genes, namely an LMX1A gene and a VAX1 gene. A urine sample is kept before the operation of a detected bladder cancer patient, and the specific CpG sites of the genes are subjected to high methylation among people who are determined to suffer from a relapse in the follow-up visit process. Thus, the two genes can be used as biological markers for the relapse of bladder cancer. The invention can be used as the basis for designing a kit for prognosing and diagnosing the relapse of bladder cancer, and is applicable to the prognosis detection of bladder cancer in hospital, the postoperative follow-up visit and the postoperative monitoring of the community health center on people subjected to bladder cancer operation.

Description

technical field [0001] The present invention relates to biomedical detection; more specifically, the present invention relates to a new biomarker (urine marker) for predicting bladder cancer recurrence, and a diagnostic reagent and kit for bladder cancer recurrence prognosis based on the marker. Background technique [0002] Transitional cell carcinoma of the bladder is the most common type of bladder tumor, and the incidence peaks at the age of 60 to 69 years. In some large cities in China, with the acceleration of industrialization and changes in personal diet, smoking and other living habits, the incidence rate has a clear upward trend , and there is a trend of younger onset. At the same time, the postoperative recurrence rate of bladder cancer is extremely high, about 50-70%, and the recurrence time is within 1 to 2 years. Some cases are accompanied by increased malignancy or enhanced infiltration ability of the tumor, which seriously affects the survival of patients. t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/113
Inventor 余坚王红阳顾健人赵仰星孙晋枫张红宇顾峻王韦何英华郭士成
Owner SHANGHAI INST OF ONCOLOGY
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