Unlock instant, AI-driven research and patent intelligence for your innovation.

Application of circular RNAs as molecular markers for colorectal cancer

A colorectal cancer and circular technology, which is applied in the field of biomedicine, can solve the problems that there are no plasma circular RNA markers for colorectal cancer, and achieve early diagnosis, reduce mortality, and wide application prospects

Active Publication Date: 2021-07-27
JINAN UNIVERSITY
View PDF0 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although there are more and more studies on circular RNA, which has become a potential biomarker for the early diagnosis of a new generation of tumors, so far, there is no plasma circular RNA marker that can be used for early diagnosis and screening of colorectal cancer related reports

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Application of circular RNAs as molecular markers for colorectal cancer
  • Application of circular RNAs as molecular markers for colorectal cancer
  • Application of circular RNAs as molecular markers for colorectal cancer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0061] Example 1: Detection of expression of hsa_circ_0005758 in colorectal cancer tissue by Real-time PCR

[0062] 1. Sample collection and preparation

[0063] 10 patients with colorectal cancer, 10 patients with colonic polyps and 10 healthy volunteers were collected. 4ml of whole blood was drawn from each, and centrifuged at 12,000×g at 4°C for 10 minutes to remove possible impurities and collect plasma samples. The collected plasma samples were stored in a -80°C refrigerator for later use.

[0064] 2. RNA extraction

[0065] The above plasma samples were dissolved on ice, and 750 μl of TRIzol LS Reagent and 20 μl of glacial acetic acid were added respectively, and the tube body was shaken vigorously by hand until mixed. After homogenization, incubate the sample at room temperature for 5 minutes to completely dissociate the nucleic acid-protein complex, add 0.2ml of chloroform, vibrate the tube body manually for 15 seconds, and incubate at room temperature for 2-3 minute...

Embodiment 2

[0080] Example 2: Effect of hsa_circ_0005758 expression on colorectal cancer cells detected by colorectal cancer cell proliferation assay

[0081] 1. Intestinal Cancer Cell Culture

[0082]Intestinal cancer cell lines HCT116 and HT29 (purchased from ATCC) are adherent cells, which were cultured in culture medium containing 10% (v / v) fetal bovine serum (McCoy's 5A (Modified) Medium, purchased from Thermo Fisher Scientific). Adjust the cell concentration to 3~5×10 6 / L, inoculated in the culture dish, the culture condition is 37 ℃, 5% (v / v) CO 2 , 95% humidity. The growth of the cells was observed every day, and the culture medium was replaced. The cells grew in a single layer adherently.

[0083] 2. hsa_circ_0005758 transfection

[0084] Intestinal cancer cells were divided into 1 × 10 5 Spread into a 24-well plate per well, and after culturing for 18-24 hours, the number of cells is about 2×10 5 / well, transfected with hsa_circ_0005758 lentivirus (purchased from Guangzho...

Embodiment 3

[0089] Example 3: Colorectal cancer cell proliferation assay detects the effect of interfering with the expression of hsa_circ_0005758 on colorectal cancer cells

[0090] 1. Intestinal Cancer Cell Culture

[0091] Intestinal cancer cell lines HCT116 and HT29 (purchased from ATCC) are adherent cells, which were cultured in culture medium containing 10% (v / v) fetal bovine serum (McCoy's 5A (Modified) Medium, purchased from Thermo Fisher Scientific). Adjust the cell concentration to 3~5×10 6 / L, inoculated in the culture dish, the culture condition is 37 ℃, 5% (v / v) CO 2 , 95% humidity. The growth of the cells was observed every day and the culture medium was replaced. The cells grew in a single layer adherent to the wall.

[0092] 2. Interfering with hsa_circ_0005758 transfection

[0093] The target sequence of interference hsa_circ_0005758 is: GCTAGCACTGGACAGTCAC. RNA that interferes with hsa_circ_0005758 was synthesized (si-hsa_circ_0005758, siRNA). Taking a six-well pla...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention discloses the application of circular RNA as a colorectal cancer molecular marker. The present invention finds for the first time that hsa_circ_0005758 is related to colorectal cancer, and the expression of hsa_circ_0005758 in the plasma of patients with colorectal cancer is reduced. Therefore, by detecting the expression of hsa_circ_0005758 in the subject, it can be judged whether the subject has colorectal cancer or whether there is a patient with Risk of colorectal cancer. The hsa_circ_0005758 of the present invention can be used as a molecular marker related to the diagnosis and treatment of colorectal cancer, which can realize the early diagnosis of colorectal cancer; it can also be used as an effective target for screening the diagnosis and treatment of colorectal cancer, and can be made into diagnostic kits, test strips, and chips and other diagnostic tools. At the same time, the present invention also proves that hsa_circ_0005758 can inhibit the proliferation of intestinal cancer cells, so it can be used in the field of antitumor drugs.

Description

technical field [0001] The invention belongs to the field of biomedicine, in particular to the application of circular RNA as a colorectal cancer molecular marker. Background technique [0002] Since the early symptoms of gastrointestinal tumors are not obvious and the specificity of screening indicators is not high, early detection and early diagnosis have become a major problem, and the accuracy and sensitivity of postoperative recurrence detection indicators are not ideal. The research of circular RNA It has brought dawn to the early diagnosis of gastrointestinal tumors. [0003] Circular RNAs belong to non-coding RNAs, which are closed RNAs formed by non-covalent linkage of exons and or introns (Guarnerio, J., et al., Oncogenic Role of Fusion-circRNAs Derived from Cancer-Associated Chromosomal Translocations. Cell, 2016.166(4):p.1055-1056.). The characteristics of circular RNA mainly include the following points: ①Circular RNA widely exists in eukaryotic organisms, wit...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6886
CPCC12Q1/6886C12Q2600/106C12Q2600/158C12Q2600/178
Inventor 施颖
Owner JINAN UNIVERSITY