Application of hsa_circ_0049154 as prostatic cancer molecular target in preparation of drug and kit

A prostate cancer and molecular target technology, which is applied in the determination/testing of microorganisms, biochemical equipment and methods, DNA/RNA fragments, etc., can solve the problems of difficult diagnosis, high misdiagnosis rate, and unusability, and achieve sensitivity, convenience, The effect of high specificity and convenient sampling

Inactive Publication Date: 2018-11-02
FUDAN UNIV
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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_0049154 as prostatic cancer molecular target in preparation of drug and kit
  • Application of hsa_circ_0049154 as prostatic cancer molecular target in preparation of drug and kit
  • Application of hsa_circ_0049154 as prostatic cancer molecular target in preparation of drug and kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] Example 1: Content detection of hsa_circ_0049154 in prostate cancer tissue samples and normal tissue samples.

[0053] The hsa_circ_0049154 detection primer sequence used in this example is:

[0054] hsa_circ_0049154 upstream primer sequence: 5'- GGGAACCAATCTGCCCTTTT-3' (SEQ ID NO.4)

[0055] The downstream primer sequence of hsa_circ_0049154: 5'- ATGGCGGTCTGATAACCACTT-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] Obtain post-operative tissue samples from patients undergoing radical prostatectomy and patients who have undergone lymphadenectomy at the same time

[0059] Excised tissue samples served as controls. Extract the total RNA from the aforementioned tissue samples into a 1.5 ml centrifuge tube free of DNA and RNase contamination.

[0060] The kit for extracting total RNA from tissue samples was purchased from Beijing Kangwei Century Biotechnology Co., Ltd. The co...

Embodiment 2

[0076] Example 2: Content detection of hsa_circ_0049154 in prostate cancer plasma samples and normal plasma samples.

[0077] The hsa_circ_0049154 detection primer sequence used in this example is:

[0078] hsa_circ_0049154 upstream primer sequence: 5'- GGGAACCAATCTGCCCTTTT-3' (SEQ ID NO.4)

[0079] The downstream primer sequence of hsa_circ_0049154: 5'- ATGGCGGTCTGATAACCACTT-3' (SEQ ID NO.5).

[0080] The main steps of this embodiment are as follows:

[0081] 1. Extraction of total RNA from plasma samples

[0082] 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;

[0083] 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 ultravio...

Embodiment 3

[0099] Example 3: Detection of hsa_circ_0049154 silencing efficiency of hsa_circ_0049154 siRNA in prostate cancer cell lines.

[0100] The siRNA sequence of the prostate cancer molecular target hsa_circ_0049154 used in this example is:

[0101] hsa_circ_0049154 siRNA sense strand sequence: 5'- GCAAGUGGUUAUCAGACCG -3' (SEQ ID NO.6)

[0102] hsa_circ_0049154 siRNA antisense strand sequence: 5'- CGGUCUGAUAACCCACUUGC-3' (SEQ ID NO.7)

[0103] The siRNA sequence of the negative control (NC) used is

[0104] NC siRNA sense strand sequence: 5'- UUCUCCGAACGUGUCACGU -3' (SEQ ID NO.8)

[0105] NC siRNA antisense strand sequence: 5'- ACGUGACACGUUCGGAGAA -3' (SEQ ID NO.9).

[0106] The main steps of this embodiment are as follows:

[0107] 1. siRNA transfection of cells (taking a six-well plate as an example)

[0108] 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 25...

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Abstract

The invention belongs to the technical field of biology and particularly relates to an application of hsa_circ_0049154 as a prostatic cancer molecular target in preparation of a drug and a kit. The nucleotide sequence of hsa_circ_0049154 is shown in SEQ ID NO.1. The application comprises an application of the prostatic cancer molecular target in screening a drug for preventing, relieving and / or treating prostatic cancer, an inhibitor of the prostatic cancer molecular target and an application of the inhibitor in preparing a drug for preventing, relieving and / or treating prostatic cancer, a prostatic cancer diagnosis kit containing the prostatic cancer molecular target as well as an 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 used for diagnosing the prostatic cancer, the molecular target and the diagnosis kit containing the molecular target have the characteristics of being simple to operate, convenient in material taking, safe, free from wound, higher in specificity and sensitivity and capable of facilitating mass 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_0049154 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): C12Q1/6886C12N15/11
CPCC12Q1/6886C12Q2600/136C12Q2600/178
Inventor 李瑶吴海孔哲万学超鲁亚莉张亚龙
Owner FUDAN UNIV
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