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Prostatic cancer molecular target RP11-1023L17.1 and application thereof to diagnostic kit

A prostate cancer and molecular target technology, applied in RP11-1023L17.1 as a prostate cancer molecular target and its application in diagnostic kits, can solve the problems of difficult diagnosis, low diagnostic coincidence rate, high misdiagnosis rate, etc., and achieve Sensitivity is convenient, operation is simple, and the effect of high specificity

Inactive Publication Date: 2015-12-16
FUDAN UNIV +1
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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, but it is already at an advanced stage, and it is not easy to cure; (3) This test cannot be used when there is a disease in the rectum; (4) Inexperienced doctors may miss or misdiagnose
Under normal circumstances, the PSA in the blood is not high (not higher than 4ng / ml). When in the state of prostate cancer and other prostate diseases, the increase of PSA has become the most sensitive tumor marker for screening prostate cancer, but it also exists Certain limitations: (1) Blood testing is required, which will cause certain damage to the patient; (2) Elevated PSA is also common with non-prostate cancer diseases, such as prostatitis, prostatic hypertrophy, etc., so it is not easy to diagnose; (3) Elevated PSA is diagnosed In the case of prostate cancer, the patients are often in the middle and late stage, and the purpose of early diagnosis cannot be achieved
Ultrasonic detection of the prostate is simple, intuitive, and non-invasive. It provides positioning by displaying the size, number, location, density, edge, shape, presence or absence of calcification, and the shape, size, number, and distribution of calcification, as well as surrounding halos and skin changes. and qualitative signs and judge the nature of the lesion; its limitations are: (1) it is easy to miss the diagnosis of dense small cancer focus; (2) sometimes it cannot provide a clear qualitative diagnosis; (3) because it cannot show the internal structure of the tumor (4) There is a high misdiagnosis rate for some solid benign and malignant masses lacking typical signs

Method used

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  • Prostatic cancer molecular target RP11-1023L17.1 and application thereof to diagnostic kit
  • Prostatic cancer molecular target RP11-1023L17.1 and application thereof to diagnostic kit
  • Prostatic cancer molecular target RP11-1023L17.1 and application thereof to diagnostic kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Example 1: Detection of RP11-1023L17.1 in prostate cancer tissue samples and normal tissue samples

[0039] The main steps of this embodiment are as follows:

[0040] 1. Extraction of total RNA from tissue samples

[0041] Postoperative tissue samples were obtained from patients undergoing radical prostatectomy, while tissue samples resected from patients undergoing lymphadenectomy were obtained as controls. Extract the total RNA from the aforementioned tissue samples into a 1.5 ml centrifuge tube free of DNA and RNase contamination.

[0042] The kit for extracting total RNA from tissue samples was purchased from Beijing Kangwei Century Biotechnology Co., Ltd. The concentration of the extracted total RNA was determined by using a ThermNanoDrop2000c spectrophotometer to measure the ratio of the 260 / 280nm ultraviolet wavelength.

[0043] 2. Quantitative detection of lncRNA in tissue samples

[0044] (1) Reverse transcribe RNA to obtain single-stranded cDNA

[0045]...

Embodiment 2

[0057] Example 2: Detection of RP11-1023L17.1 silencing efficiency by RP11-1023L17.1-siRNA in prostate cancer cell lines

[0058] The siRNA sequence of the prostate cancer molecular target RP11-1023L17.1 used in this example is:

[0059] RP11-1023L17.1 siRNA sense strand sequence: 5'-GCGUCUAAGAGAGUACUUU-3' (SEQ ID NO.6)

[0060] RP11-1023L17.1 siRNA antisense strand sequence: 5'-AAAGUACUCUCUUAGACGC-3' (SEQ ID NO.7).

[0061] The siRNA sequence of the negative control (NC) used was:

[0062] NCsiRNA sense strand sequence: 5'-UUCUCCGAACGUGUCACGU-3' (SEQ ID NO.8)

[0063] NCsiRNA antisense strand sequence: 5'-ACGUGACACGUUCGGAGAA-3' (SEQ ID NO.9).

[0064] The main steps of this embodiment are as follows:

[0065] 1. siRNA transfection of cells (taking a six-well plate as an example)

[0066] The prostate cancer PC-3 cell line was inoculated into a six-well plate, so that the next day the cell confluence was about 40-50%, and 5 μL of Lipofectamine2000 and 250 μL of Opti-MEM...

Embodiment 3

[0073] Example 3: The RP11-1023L17.1-siRNA significantly inhibits the proliferation of prostate cancer cells and can be used for the preparation of prostate cancer drugs.

[0074] The main steps of this embodiment are as follows:

[0075] 1. siRNA transfection of cells (taking a six-well plate as an example)

[0076] The transfection method was the same as in Example 2.

[0077] 2. Cell proliferation experiment

[0078] After the cells were digested, they were resuspended, seeded in 96 wells, and a control well with only 100 μL of 1640 medium was set. After 0h, 24h, and 48h of inoculation, add 10ul of CKK-8 reagent to each well, and continue to incubate for 2 hours in the cell culture incubator. Use the optical absorption value (OD) of each well at the detection wavelength of 450nm in a microplate reader, and the reference wavelength is 630nm (600-650nm). Take the average value of the light absorption value of each well minus the light absorption value of the blank well ...

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Abstract

The invention belongs to the biotechnology field and in particular provides a prostatic cancer molecular target RP11-1023L17.1 and an application thereof to a diagnostic kit. RP11-1023L17.1 is IncRNA (long non-coding ribonucleic acid). A nucleotide sequence of RP11-1023L17.1 is shown in SEQ ID NO.1. The invention comprises an application of the prostatic cancer molecular target to screening drugs for preventing, relieving and / or treating prostatic cancers, an inhibitor of the prostatic cancer molecular target, an application of the inhibitor to preparation of drugs for preventing, relieving and / or treating prostatic cancers, the prostatic cancer diagnostic kit containing the prostatic cancer molecular target and applications of the prostatic cancer molecular target and the diagnostic kit to screening, diagnosis, treatment, status monitoring and prognostic monitoring of prostatic cancer high risk groups. Being used for diagnosing prostatic cancers, the molecular target and the diagnostic kit containing the molecular target are simple to operate and convenient to draw, are safe and non-invasive and have the characteristics of higher specificity and sensitivity and easiness in mass screening.

Description

technical field [0001] The invention belongs to the field of biotechnology, and specifically relates to a prostate cancer molecular target RP11-1023L17.1 and its application in a diagnostic kit, including the screening of high-risk groups of prostate cancer, the diagnosis of prostate cancer, and the diagnosis of prostate cancer through the biomolecular target. Applications in the fields of monitoring of cancer treatment and status, monitoring of prostate cancer-guided medication, and detection of prostate cancer prognosis. Background technique [0002] According to the relevant content in the "ASCO Annual Report: 2015 Clinical Oncology Progress", liquid biopsy technology is listed as one of the next decade trends in the field of tumor treatment. Some molecular markers in the blood have been used in the diagnosis of some diseases, such as alanine aminotransferase for the diagnosis of hepatitis, and the number of white blood cells for judging inflammation. Circulating tumor D...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/113A61K48/00A61P35/00C12Q1/68C12N15/11
Inventor 李瑶鲍升林黄文华万学超张亚龙吴海
Owner FUDAN UNIV
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