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Application of hsa_circ_0012755 as prostatic cancer molecular target in preparing medicine and kit

A prostate cancer and molecular target technology, which is applied in drug combinations, biochemical equipment and methods, and microbial measurement/inspection, can solve the problems of difficult diagnosis, low diagnostic coincidence rate, high misdiagnosis rate, etc., and achieve sensitivity, convenience, Simple operation, high specific effect

Active Publication Date: 2018-10-09
FUDAN UNIV
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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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  • Application of hsa_circ_0012755 as prostatic cancer molecular target in preparing medicine and kit
  • Application of hsa_circ_0012755 as prostatic cancer molecular target in preparing medicine and kit
  • Application of hsa_circ_0012755 as prostatic cancer molecular target in preparing medicine and kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] Example 1: Content detection of hsa_circ_0012755 in prostate cancer tissue samples and normal tissue samples.

[0053] The hsa_circ_0012755 detection primer sequence used in this example is:

[0054] hsa_circ_0012755 upstream primer sequence: 5'- GGTTTCAGCAGCCCCTCC -3' (SEQ ID NO.4)

[0055] The upstream primer sequence of hsa_circ_0012755: 5'-CCTTAATGTCAGCGTCACTTGG-3' (SEQ ID NO.5).

[0056] The main steps of this embodiment are as follows:

[0057] 1. Extraction of total RNA from tissue samples

[0058] 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 of the aforementioned tissue samples in a 1.5 ml centrifuge tube without DNA and RNase contamination;

[0059] The kit for extracting total RNA from tissue samples was purchased from Beijing Kangwei Century Biotechnology Co., Ltd. The extracted total RNA is...

Embodiment 2

[0074] Example 2: Content detection of hsa_circ_0012755 in prostate cancer plasma samples and normal plasma samples.

[0075] The hsa_circ_0012755 detection primer sequence used in this example is:

[0076] hsa_circ_0012755 upstream primer sequence: 5'- GGTTTCAGCAGCCCCTCC -3' (SEQ ID NO.4)

[0077] The upstream primer sequence of hsa_circ_0012755: 5'-CCTTAATGTCAGCGTCACTTGG-3' (SEQ ID NO.5).

[0078] The main steps of this embodiment are as follows:

[0079] 1. Extraction of total RNA from plasma samples

[0080] Plasma samples from 26 patients with prostate cancer and 19 normal subjects were obtained as controls. Extract the total RNA of the aforementioned tissue samples in a 1.5 ml centrifuge tube without DNA and RNase contamination;

[0081] A kit for extracting total RNA from plasma samples was purchased from QIAGEN. The extracted total RNA is determined by using a ThermNanoDrop2000c spectrophotometer to determine the concentration of the ratio of 260 / 280nm ultraviolet...

Embodiment 3

[0096] Example 3: Detection of hsa_circ_0012755 silencing efficiency of hsa_circ_0012755 siRNA in prostate cancer cell lines.

[0097] The siRNA sequence of the prostate cancer molecular target hsa_circ_0012755 used in this example is:

[0098] Sequence of hsa_circ_0012755 siRNA sense strand: 5'- CCCAGAGAUUCAAGUCAAU -3' (SEQ ID NO.6)

[0099] hsa_circ_0012755 siRNA antisense strand sequence: 5'- AUUGACUUGAAUCUCUGGG -3' (SEQ ID NO.7)

[0100] The siRNA sequence of the negative control (NC) used is

[0101] NC siRNA sense strand sequence: 5'- UUCUCCGAACGUGUCACGU -3' (SEQ ID NO.8)

[0102] NC siRNA antisense strand sequence: 5'- ACGUGACACGUUCGGAGAA -3' (SEQ ID NO.9).

[0103] The main steps of this embodiment are as follows:

[0104] 1. siRNA transfection of cells (taking a six-well plate as an example)

[0105] The prostate cancer PC-3 cell line was inoculated into a six-well plate, and the cell density reached about 60-70% two days after plating, and 5 μL of Lipofectamine ...

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Abstract

The invention belongs to the technical field of biologics and particularly discloses the application of hsa_circ_0012755 as a prostatic cancer molecular target in preparing a medicine and a kit. hsa_circ_0012755 has a nucleotide sequence of SEQ ID NO.1 as shown in the specification. The invention discloses the application of the prostatic cancer molecular target in preparing medicines for screening, preventing, relieving and / or treating prostatic cancer, an inhibitor with the prostatic cancer molecular target, the application of the inhibitor in preparing medicines for preventing, relieving and / or treating prostatic cancer, a prostatic cancer diagnosis kit with the prostatic cancer molecular target, and the application of the prostatic cancer molecular target and the diagnosis kit in screening, diagnosis, treatment, situation monitoring and prognosis monitoring of high risk groups of prostatic cancer. When being adopted for prostatic cancer diagnosis, the molecular target and the diagnosis kit with the molecular target are simple to operate, convenient in material obtaining and safe and noninvasive, and in addition have the characteristics of being high in specificity and sensitivity and easy in large-scale screening.

Description

technical field [0001] The invention belongs to the field of biotechnology, and specifically relates to the application of a prostate cancer molecular target hsa_circ_0012755 in the preparation of medicines and diagnostic kits, including the screening of high-risk groups of prostate cancer, the diagnosis of prostate cancer, the treatment of prostate cancer and the The monitoring of the status of prostate cancer, the monitoring of prostate cancer guidance medication and the application of prostate cancer prognosis detection and other fields. 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. C...

Claims

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Application Information

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IPC IPC(8): C12Q1/6886C12Q1/6851C12N15/11A61K45/00A61P35/00
CPCA61K45/00A61P35/00C12Q1/6851C12Q1/6886C12Q2600/178C12Q2600/166C12Q2600/136C12Q2600/158C12Q2600/118C12Q2531/113C12Q2563/107C12Q2545/101
Inventor 李瑶吴海孔哲万学超鲁亚莉张亚龙
Owner FUDAN UNIV
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