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Prostatic cancer circRNA marker and application thereof

A prostate cancer and marker technology, applied in the field of biomedicine, can solve the problems of low specificity and sensitivity, achieve high specificity and sensitivity, long storage time, high detection sensitivity and specificity

Active Publication Date: 2021-01-15
ZHUJIANG HOSPITAL SOUTHERN MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Prostate-specific antigen PSA, Gleason score, pathological stage and other indicators have low specificity and sensitivity in predicting postoperative biochemical recurrence. Therefore, there is an urgent need for more accurate biomarkers for predicting postoperative biochemical recurrence. In this way, the recurrence risk of patients can be refined and stratified, and high-risk patients can be accurately provided with adjuvant therapy to improve the prognosis

Method used

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  • Prostatic cancer circRNA marker and application thereof
  • Prostatic cancer circRNA marker and application thereof
  • Prostatic cancer circRNA marker and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1, screening of prostate cancer circRNA markers

[0029] 1. Source and acquisition of circRNA data

[0030] The circRNA sequencing data and corresponding clinical information used in the present invention come from GSE113124 in the Gene Expression Omnibus (GEO) (http: / / www.ncbi.nlm.nih.gov / geo / ) database, which includes 144 radical prostatectomy patients later patients.

[0031] 2. Screening of circRNA markers in prostate cancer

[0032]In the circRNA expression matrix, 546 target circRNAs were selected with FPKM>1 as the standard. Prostate cancer biochemical recurrence was used as an indicator to construct a circRNA prediction model. Logistic regression analysis was performed on 546 circRNAs using the 'survival' and 'survminer' R packages, and 28 groups of circRNAs with p<0.05 were obtained. Using Lasso cox regression and cross-validation to further reduce the dimensionality of 28 groups of circRNAs, and finally obtained 8 groups of circRNAs. Draw the Kapl...

Embodiment 2

[0040] Example 2, detection of prostate cancer circRNA marker primer design and kit design

[0041] In this example, 8 groups of prostate cancer circRNA markers (including circ_30029, circ_117300, circ_176436, circ_112897, circ_178252, circ_115617, circ_14736 and circ_17720) screened in Example 1 were used.

[0042] 本实施例中采用circPrimer引物设计软件设计,分别根据circBase及CIRCpediav2提供的序列:hsa_circ_0005882、hsa_circ_0001747、hsa_circ_0078607、HSA_CIRCpedia_130167、hsa_circ_0084615、hsa_circ_0007324、hsa_circ_0000072、hsa_circ_0009117、内参选用GAPDH,引物设计后由天一辉远公司合成 .

[0043] ① When the prostate cancer circRNA marker is circ_30029, the primers for specifically amplifying circ_30029 (as shown in SEQ ID NO.1 and SEQ ID NO.2) are as follows:

[0044] Forward primer: TCTCTGTGCACGACTCTCAG;

[0045] Reverse primer: TGGCTCTTTGGGCTATGTCT; the amplification length is 201bp.

[0046] ② When the prostate cancer circRNA marker is circ_117300, the primers for specifically amplifying circ_117300 (as shown in SEQ ID NO.3 ...

Embodiment 3

[0070] Example 3, Extraction of tissue sample RNA to be tested, reverse transcription PCR and timed quantitative PCR

[0071] 1. Extract RNA from tissue samples to be tested

[0072] 1) Take fresh tissue, cut into small pieces (50-100mg;)

[0073] 2) Put the tissue block into a mortar filled with liquid nitrogen;

[0074] 3) Grind the tissue with a pestle to make it into powder;

[0075] 4) After the liquid nitrogen volatilizes during the grinding process, it needs to be replenished in time to keep the sample in a frozen state to prevent RNA degradation;

[0076] 5) Transfer the ground tissue powder to an RNase free centrifuge tube, add an appropriate volume of Trizol at a ratio of 50-100 mg tissue / ml Trizol, and control the tissue volume to no more than 10% of the Trizol volume;

[0077] 6) Place at room temperature for 10 minutes to fully lyse (or transfer to -80°C for long-term storage);

[0078] 7) Add chloroform to 200 μl / ml Trizol, shake vigorously for 30 seconds, and ...

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Abstract

The invention relates to the technical field of biomedicine, in particular to a prostatic cancer circRNA marker and application thereof. The prostatic cancer circRNA marker is circ_30029, circ_117300,circ_176436, circ_112897, circ_178252, circ_115617, circ_14736 and circ_17720. The prostatic cancer circRNA marker disclosed by the invention has very high specificity and sensitivity, and the prostatic cancer circRNA marker has expression difference in prostatic cancer tissues and para-carcinoma tissues of a patient, so that the prostatic cancer circRNA marker can be used as a novel biomarker for detecting prostatic cancer, and a new reference index can also be provided for judging the biochemical recurrence risk of the prostatic cancer patient after a radical operation.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to a prostate cancer circRNA marker and its application. Background technique [0002] Prostate cancer (Prostatic Cancer, PCa) is a common malignant tumor, its morbidity and mortality peak around 70 years old, accounting for the second in the global tumor incidence and the sixth in the mortality. In 2016, there were approximately 180,890 new cases and 26,120 deaths in the United States. However, prostate cancer has significant racial disparities. In the United States alone, the incidence of prostate cancer in African-Americans is twice that of Caucasians; in addition, its incidence in Asia is also significantly lower than in Western countries. Therefore, intra-ethnic genomic analysis of prostate cancer is urgently needed. [0003] Prostate cancer has strong individual variability, and different patients respond to the same treatment plan very differently. Some patients live ...

Claims

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Application Information

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IPC IPC(8): C12Q1/6886
CPCC12Q1/6886C12Q2600/158C12Q2600/178C12Q2600/118
Inventor 卢剑铭毛向明王烁
Owner ZHUJIANG HOSPITAL SOUTHERN MEDICAL UNIV
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