Application of circular RNAcirc0001610 and expression product thereof in medicines for diagnosing and treating bladder cancer

A technology for expressing products and therapeutic drugs, applied in the field of biomedicine, can solve the problems of high recurrence and high metastasis rate of bladder cancer and lack of control methods

Active Publication Date: 2021-08-20
清远市人民医院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although localized bladder tumors can be controlled by surgical resection, there is still a lack of effective control methods for the high recurrence and high metastasis rates of bladder cancer after surgery

Method used

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  • Application of circular RNAcirc0001610 and expression product thereof in medicines for diagnosing and treating bladder cancer
  • Application of circular RNAcirc0001610 and expression product thereof in medicines for diagnosing and treating bladder cancer
  • Application of circular RNAcirc0001610 and expression product thereof in medicines for diagnosing and treating bladder cancer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Example 1, RT-PCR reaction detection of circular RNAcirc0001610 expression in bladder cancer tissue

[0035] The specific experimental plan is as follows:

[0036] 1. RNA extraction

[0037] 1) Tissue processing: Add about 10 mg of bladder cancer tissue to 1 ml Trizol, homogenize with a homogenizer; centrifuge at 12000 g for 15 minutes, and take the supernatant.

[0038]2) Add 200ul chloroform to the supernatant, mix vigorously up and down for half a minute, and let stand for 3 minutes.

[0039] 3) Place the supernatant after standing at 4°C and centrifuge at 12000g for 15 minutes. At this time, it can be seen that the lysate is divided into three layers: the upper layer is RNA in the water phase; the middle layer is DNA, lipids, etc.; the lower layer is cell residues, proteins, Polysaccharides etc.

[0040] 5) Take the supernatant into a new EP tube; add an equal volume of isopropanol, mix well, let stand for 10 minutes, then place at 4°C and centrifuge at 12000g ...

Embodiment 2

[0055] Example 2, QPCR detection of expression of circular RNA Circ0001610 in bladder cancer tissue

[0056] 1. RNA extraction: same as Example 1;

[0057] 2. cDNA reverse transcription: same as Example 1;

[0058] 3. QPCR amplification experiment

[0059] Experimental system:

[0060]

[0061]

[0062] 1) Reaction conditions:

[0063] Step 1: 95°C for 2 minutes;

[0064] The second step (40 cycles): 95°C for 3 seconds, 60°C for 30 seconds;

[0065] The third step 60-95 ℃ melting curve.

[0066] 2) Amplify the target gene on the machine

[0067] 3) qPCR relative quantitative results

[0068] The formula for calculating the relative expression of the target gene is: 2-△△Ct=2-[(△Ct)Test-(△Ct)Control]. Ct object is the Ct value of the target gene, and Ct housekeeper is the Ct value of the housekeeping gene. △Ct=Ct Purpose-Ct Housekeeper, indicating the relative Ct value of the target gene of each sample relative to the housekeeping gene, △△Ct=(△Ct)Test-(△Ct)Contr...

Embodiment 3

[0070] Example 3, the effect of siRNA interference on the expression of circular RNA Circ0001610 on the proliferation of bladder cancer cells

[0071] Three specific siRNAs for knocking down circular RNA Circ0001610 were constructed against the circularization site of circular RNA Circ0001610. The specific siRNA is synthesized by Suzhou Jima Gene, and the specific siRNA includes siRNA-1, siRNA-2 and siRNA-3, the siRNA-1 is composed of sequences such as SEQ ID NO:10 and SEQ ID NO:11, and the siRNA- 2 consists of sequences such as SEQ ID NO: 12 and SEQ ID NO: 13, and the siRNA-3 consists of sequences such as SEQ ID NO: 14 and SEQ ID NO: 15. The three siRNAs and the negative control fragment (NC) were transfected in 5637 and J82 cells, respectively, and the cells were collected 48 hours after transfection to extract RNA for qPCR detection, and the relative quantification of the circular RNA Circ0001610 of the cells was performed.

[0072] Table 2

[0073]

[0074]

[00...

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Abstract

The invention relates to the technical field of biological medicines, and particularly discloses application of a circular RNAcirc0001610 and an expression product thereof in medicines for diagnosing and treating bladder cancer, and the nucleotide sequence of the circular RNAcirc0001610 is as shown in SEQ ID NO: 1. Experiments show that the expression of the circular RNAcirc0001610 and the expression product thereof in bladder cancer tissues is obviously up-regulated. In human bladder cancer cells 5637 and bladder cancer cells J82, the proliferation, migration and invasion levels of the bladder cancer cells are remarkably inhibited by knocking down the expression level of the circular RNAcirc0001610, that is, the growth of bladder cancer cell strains can be inhibited by down-regulating the expression of the circular RNAcirc0001610.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to the application of a circular RNA circ0001610 and its expression product in diagnosing and treating bladder cancer. Background technique [0002] Bladder cancer is the most common malignant tumor of the urinary system, accounting for the first place in the incidence of genitourinary system tumors in my country. According to the National Cancer Survey, there were 80,000 new cases of bladder cancer in 2015, ranking 11th in the incidence of new malignant tumors. Although localized bladder tumors can be controlled by surgical resection, there is still a lack of effective control methods for the high recurrence and high metastasis rates of bladder cancer after surgery. Therefore, elucidating the molecular mechanism of the occurrence and development of bladder cancer, finding new targets for regulating the invasion and metastasis of bladder cancer, and developing targeted anti-tum...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886C12N15/113A61K45/00A61K31/713A61P35/00
CPCC12Q1/6886C12N15/113A61K45/00A61P35/00C12Q2600/158C12Q2600/178C12Q2600/136C12N2310/14C12N2310/113
Inventor 张钰莹曾健文朱宝益朱思华
Owner 清远市人民医院
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