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Application of lncRNA IGFL2-AS1 as a diagnostic marker for colon cancer

A technique for colon cancer, biological samples, applied in the field of tumor molecular biology

Active Publication Date: 2022-07-12
西安奥锐精创生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, there are few reports on the role of lncRNA in the development of colon cancer.

Method used

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  • Application of lncRNA IGFL2-AS1 as a diagnostic marker for colon cancer
  • Application of lncRNA IGFL2-AS1 as a diagnostic marker for colon cancer
  • Application of lncRNA IGFL2-AS1 as a diagnostic marker for colon cancer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1: Real-time PCR detection of lncRNA IGFL2-AS1

[0032] Extract total RNA:

[0033] From colon cancer tissue samples and adjacent tissue samples, add 1 mL After the Reagent was placed at room temperature (15-30°C), the nucleoprotein complexes were completely separated. After 5 min, add 0.2 mL of chloroform, cover the tube, shake vigorously by hand for 15 s, and leave at room temperature for 2-3 min. Centrifuge at 12 000 g for 15 min at 2-8 °C, and transfer the upper colorless aqueous phase to another 1.5 mL Eppendorf tube. Add 0.5 mL of isopropanol and leave at room temperature for 10 min. Centrifuge at 12 000 g for 10 min at 2-8 °C and discard the supernatant. The RNA pellet was washed with 1 mL of 75% ethanol and mixed with a vortex mixer. Centrifuge at 7500g for 5 min at 2-8°C and discard the supernatant. Use RNase-free ddH after air drying for 5-10 min 2 O dissolves RNA, blows and sucks repeatedly with a micropipette and incubates at 55-60 °C for 10 ...

Embodiment example 2

[0044] Example 2: Knockdown of lncRNA IGFL2-AS1 in cancer cells

[0045] The SW620 cell line was selected for the knockdown experiment of lncRNA IGFL2-AS1, and the related gene expression and cell migration ability after knockdown were detected.

[0046] Knockdown of lncRNA IGFL2-AS1:

[0047] The knockdown of lncRNA IGFL2-AS1 was achieved by siRNA. The specific method was as follows: cells were plated on a six-well plate, and when the plate was 50% full the next day, starve with Opti-MEM for 30 minutes;

[0048] Dissolve 2.5 nmol siRNA / NC with 125 μL of DEPC water, and the amount of each well is 10 μL. Take two tubes labeled A and B, add 100 μL Opti-MEM to each tube; add 10 μL lipofectamine 3000 to tube A, add 10 μL siRNA or NC to tube B, mix well and let stand at room temperature for 5 min. The two tubes of AB were fully mixed, and added to the six-well plate for 15 min, and transfected for 6-8 h.

[0049] The siRNA used in the research of the present invention, the seque...

Embodiment example 3

[0057] Implementation Case 3: CCK-8 Experiment

[0058] Take 1×10 5 SW620 cells were seeded in a 12-well plate at 37°C with 5% CO. 2 Culture, when the cells grow to 70% confluence the next day, remove the original medium, add cells, trypsinize after 24 h, and inoculate 3 000 cells / well in a 96-well plate, with 5 duplicate wells in each group . After culturing for 1-7 days, 10 μl of CCK-8 was added to each well, and the culture was continued for 1 h. The blank well was used to adjust zero, the wavelength was 450 nm, and the OD value was detected by an automatic enzyme label reader.

[0059] The result is as image 3 shown, the cells transfected with siRNA had reduced proliferative capacity compared to tumor cells that were not knocked down.

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Abstract

The invention discloses an application of lncRNA IGFL2-AS1 as a diagnostic marker for colon cancer, and belongs to the field of tumor molecular biology. The lncRNA IGFL2-AS1 provided by the present invention is highly expressed in colon cancer, and plays an important role in the malignant transformation and tumor formation of colon cancer cells. Artificial knockdown of lncRNA IGFL2-AS1 in colon cancer cells can significantly inhibit the proliferation, migration, invasion ability and malignancy of SW620 cells, indicating that lncRNA IGFL2-AS1 plays an important role in the pathogenesis of colon cancer and can be used as a diagnostic tool for colon cancer. New molecular markers and drug targets.

Description

technical field [0001] The invention belongs to the field of tumor molecular biology, in particular to the application of a lncRNA IGFL2-AS1 as a diagnostic marker for colon cancer. Background technique [0002] Currently, colorectal cancer ranks third in the incidence and second in mortality among malignant tumors worldwide, and is considered to be a major threat to public health. Due to poor diet and living habits, the morbidity and mortality of colon cancer show a significant upward trend, and it is one of the most common gastrointestinal malignancies. In my country, there are about 170,000 new cases of colon cancer every year, and the age of onset of patients is relatively young, which has caused great harm to the health of the Chinese people. Colorectal carcinogenesis is a multistep biological process involving the dysregulation of multiple oncogenes and tumor suppressor genes. Despite a better understanding of the disease, advances in diagnostic techniques, and incre...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6886C12N15/113A61P35/00
CPCC12Q1/6886C12N15/113A61P35/00C12Q2600/158C12Q2600/178C12N2310/113C12N2310/14
Inventor 李小俊毕焕京闫飞李倩
Owner 西安奥锐精创生物科技有限公司
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