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Product for diagnosing prostate cancer and use thereof

A prostate cancer and product technology, applied in the field of genetic engineering, can solve problems such as missed diagnosis or misdiagnosis, easy missed diagnosis, and unobvious enlargement of prostate cancer

Inactive Publication Date: 2017-02-15
北京致成生物医学科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For example, the limitations of digital rectal examination mainly lie in four aspects: (1) when the prostate mass is not large, it is easy to miss the diagnosis; (2) some patients have no obvious prostate cancer enlargement, but it is already at an advanced stage, and it is difficult to cure; (3) This test cannot be used when there is a disease in the rectum; (4) Inexperienced doctors may miss or misdiagnose

Method used

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  • Product for diagnosing prostate cancer and use thereof
  • Product for diagnosing prostate cancer and use thereof
  • Product for diagnosing prostate cancer and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1 High-throughput sequencing screening of differentially expressed genes

[0033] 1. Sampling

[0034] From October 2012 to December 2015, 27 tissue samples were obtained from the Department of Urology at Peking Union Medical College Hospital. All samples were confirmed by pathological examination, including 8 cases of paracancerous tissue samples and 19 cases of prostate cancer samples. Store in -80°C low temperature refrigerator.

[0035] 2. Total RNA extraction from tissue samples

[0036] use Reagent (invitrogen, Cat. No. 15596-018) was used to extract sample RNA, and the experimental operation was carried out according to the product manual. The specific operation was as follows:

[0037] After collecting the samples, freeze them in liquid nitrogen. After taking them out, put the tissue into a pre-cooled mortar for grinding. After the tissue sample is powdered:

[0038] ① Add Trizol and store at room temperature for 5 minutes;

[0039] ② Add 0.2 mL of ...

Embodiment 2

[0048] Example 2 RT-PCR verification of the expression of FAM71F2 in prostate cancer tissues and adjacent tissues

[0049] 1. Materials

[0050] 34 samples of prostate cancer tissue and 8 samples of paracancerous tissue were selected and grouped and numbered. All samples were confirmed by pathological examination.

[0051] 2. Method

[0052] 2.1 Extract the total RNA from the tissue sample, the same as the extraction method in Example 1.

[0053] 2.2 Synthesis of cDNA by reverse transcription

[0054] use III Reverse Transcriptase (invitrogen, Cat. No. 18080-044) was used for cDNA reverse transcription, and the experimental operation was carried out according to the product manual. The specific operation was as follows:

[0055] Using a reverse transcription kit, 1 μg of total RNA was reverse-transcribed with reverse transcription buffer to synthesize cDNA. A 25 μL reaction system was used, and 1 μg of total RNA was used as template RNA for each sample. The obtained cD...

Embodiment 3

[0075] Example 3 Prostate Cancer Detection Kit

[0076] Based on the primer set obtained in Example 2, assemble the test kit for prostate cancer according to the present invention, the kit includes a primer pair for specific amplification of FAM71F2 mRNA as shown in SEQ ID NO: 1 and SEQ ID NO: 2, The primer pair for specifically amplifying the internal reference gene (GAPDH) is shown in SEQ ID NO: 3 and SEQ ID NO: 4; it also includes a SYBR Green polymerase chain reaction system, such as PCR buffer, SYBR Green fluorescent dye, and dNTPs. The composition of described PCR buffer solution is 25mMKCL, 2.5mM MgCL 2 , 200mM (NH 4 ) 2 SO 4 .

[0077] By optimizing the primer concentration and annealing temperature, the optimal reaction system was determined as shown in Table 3:

[0078] Table 3 PCR reaction system

[0079] components Amount added SYBR Green polymerase chain reaction system 12.5μL Upstream primer (10μM) 0.5μL Downstream primer (10μ...

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Abstract

The invention discloses a product for diagnosing prostate cancer. The product can diagnose prostate cancer through detecting expression levels of FAM71F2 mRNA in a biological sample or an FAM71F2 mRNA expression protein and its fragment, analog and derivatives. The invention further discloses a use of the product in the diagnosis of prostate cancer. The invention also discloses a method for detecting if the expression level of prostate cancer-related mRNA is reduced. In detection of prostate cancer, the mRNA can quickly and effectively carry out early detection and provide a therapeutic target and an important basis for clinical use such as gene therapy and drug therapy.

Description

technical field [0001] The invention relates to the field of genetic engineering, in particular to the use of FAM71F2 related to prostate cancer as a product for detecting prostate cancer and its application. Background technique [0002] Prostate cancer is one of the common male reproductive system malignancies. Worldwide, the incidence of prostate cancer ranks second among all malignant tumors in men. In the United States, the incidence of prostate cancer has surpassed that of lung cancer, becoming the first tumor that endangers men's health. The incidence of prostate cancer in Asia is far lower than that in European and American countries, but it has shown an upward trend in recent years, and the growth rate is faster than that in developed countries in Europe and America. Prostate cancer patients are mainly elderly men, usually after the age of 50, 95% occur in elderly men over 60 years old, and the incidence rate continues to increase with age. Prostate cancer often h...

Claims

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Application Information

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IPC IPC(8): C12Q1/68G01N33/68G01N33/574
CPCC12Q1/6886C12Q2600/158C12Q2600/178G01N33/57434G01N33/68
Inventor 刘昊
Owner 北京致成生物医学科技有限公司
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