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Circ-CRIM1 serving as ovarian cancer diagnosis marker and application thereof

A technology of CRIM1 and circ-crim1, which is applied in the field of molecular biology and tumor marker medicine, can solve the problems of lack of Circ-CRIM1 and achieve the effect of inhibiting growth

Active Publication Date: 2021-06-08
THE THIRD AFFILIATED HOSPITAL OF GUANGZHOU MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no relevant report on the expression of Circ-CRIM1 in ovarian cancer tissues, and finding effective early onset biomarkers and related molecular mechanisms is of great significance for guiding the clinical diagnosis, treatment and prognosis of ovarian cancer

Method used

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  • Circ-CRIM1 serving as ovarian cancer diagnosis marker and application thereof
  • Circ-CRIM1 serving as ovarian cancer diagnosis marker and application thereof
  • Circ-CRIM1 serving as ovarian cancer diagnosis marker and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Example 1 Identification of circ-CRIM1 and its detection in ovarian cancer tissues and cells.

[0026] Identification and detection of corresponding circular RNAs in human ovarian cancer cells. Total RNA was extracted from ovarian cancer tissues using TRIzol reagent (Japan, Shiga, Takara). First, add an appropriate amount of chloroform to TRIzol. After centrifugation, add the upper aqueous phase to a new centrifuge tube and add the same volume of isopropanol to precipitate the RNA. After centrifugation again, discard the supernatant, and wash the RNA pellet with 75% ethanol. The precipitate was then dried and dissolved in diethylpyrocarbonate (DEPC)-treated water. The absorbance (OD) at 260 nm was then measured using a UV spectrophotometer (Unico, Shanghai, China) to calculate the RNA concentration. Then, 2 μg of RNA was reverse transcribed into complementary DNA (cDNA) using avian myeloblastosis virus transcriptase and random primers (Japan, Shiga, Takara). Next, ...

Embodiment 2

[0033] Example 2 The level of circ-CRIM1 directly affects the tumor growth and metastasis of ovarian cancer.

[0034] 1. Construction of stable cell lines.

[0035]The overexpression plasmid (pHBLV-circ-CRIM1) and circ-CRIM1 short hairpin RNA (shRNA) plasmid were constructed by Shanghai Hanbio, and Lipofectamine 3000 (Invitrogen, Carlsbad, USA) was used for transfection. Doses of all reagents were based on manufacturer's instructions. Circ-CRIM1 expression was upregulated using circ-CRIM1 plasmid and circ-CRIM1 was silenced in OC cell lines using circ-CRIM1 short hairpin RNA (shRNA) plasmid. Puromycin (2 μg / ml) was used to select stable clonal cell lines. The sequences of circ-CRIM1 and circ-CRIM1 shRNA are shown in the sequence diagram. Subculture was performed when the cell plating rate reached 90%. At the same time, 5 μg / mL puromycin was added to the medium to select stably transfected cells. The cell density should not exceed 50% during screening. Screen for at least 2...

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Abstract

The invention relates to the technical field of molecular biology and tumor marker medicine, in particular to circ-CRIM1 serving as an ovarian cancer diagnosis marker and application thereof. The biomarker for ovarian cancer diagnosis is circular RNA CRIM (circ-CRIM1), the nucleotide sequence of the circular RNA CRIM (circ-CRIM1) is shown as SEQ ID No.1, occurrence and development of ovarian cancer can be clearly represented, and the biomarker shows a specific high expression phenomenon in human clinical ovarian cancer tissue compared with corresponding normal ovarian tissue and also shows a specific high expression phenomenon in a human ovarian cancer cell line. The invention further discloses application of the ovarian cancer diagnosis biomarker circular circ-CRIM1 in preparation of ovarian cancer diagnosis products. The circ-CRIM1 serves as the ovarian cancer diagnosis marker, provides a molecular target for ovarian cancer treatment and can be applied to preparation of anti-tumor drugs.

Description

technical field [0001] The invention relates to the technical fields of molecular biology and tumor marker medicine, in particular to circ-CRIM1 as a diagnostic marker for ovarian cancer and its application. Background technique [0002] Ovarian cancer (OC) is a relatively common malignancy in the female reproductive system and has the highest mortality rate among gynecological malignancies. Many types of OC are known, of which epithelial ovarian cancer (EOC) is the most common. EOC has no obvious symptoms in the early stage, and most patients are in the advanced stage when they are initially diagnosed, the treatment effect is poor, and the 5-year survival rate is extremely low. Clinically, if OC can be detected and diagnosed early, and comprehensive treatment methods such as radical surgery or targeted molecular drug therapy can be performed in a timely and reasonable manner, the survival rate, quality of life and survival time of OC patients can be greatly improved. Ther...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886C12N15/113
CPCC12Q1/6886C12Q2600/158C12Q2600/178
Inventor 赵杨宗志红杜玉平
Owner THE THIRD AFFILIATED HOSPITAL OF GUANGZHOU MEDICAL UNIVERSITY
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