TGF beta2 gene, TGF beta 2 polypeptide and application thereof

A gene, gene encoding technology, applied in the field of biomedicine

Inactive Publication Date: 2016-04-13
CANCER CENT OF GUANGZHOU MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] In addition, there is no report on TGFβ2 as a serological detection marker for breast cancer

Method used

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  • TGF beta2 gene, TGF beta 2 polypeptide and application thereof
  • TGF beta2 gene, TGF beta 2 polypeptide and application thereof
  • TGF beta2 gene, TGF beta 2 polypeptide and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1: Cell Culture

[0027] Human breast cancer cell lines MCF7, T47D (LuminalA type), BT474 (LuminalB type),

[0028] SKBR3 (HER2 overexpression type), MDA-MB-231, BT549, Hs578T (TNBC) are preserved by Cancer Institute of Guangzhou Medical University. The cells were cultured with RPMI-1640 containing 10% fetal bovine serum in a 37°C, 5% CO2 incubator.

Embodiment 2

[0029] Example 2: Real-time quantitative PCR analysis of the expression of TGFβ2 in various breast cancer cell lines

[0030] Various cultured cells grown to logarithmic phase were taken, and total RNA was extracted by TRIZOL method. After DNaseI digestion, measure the RNA. Use oligdT as a primer to take an equal amount of RNA and reverse transcribe it into cDNA according to the ReverseTranscriptionSystem program of Fermentas Company. Using cDNA as a template, according to the SYBGreen operating instructions, use the following primers for real-time quantitative PCR amplification, primers for the coding region of the TGFβ2 gene:

[0031] SEQ ID NO: 3 Forward: 5'-TAGACATGCCGCCCTTCTTC-3'

[0032] SEQ ID NO: 4 Reverse: 5'-CAAGGTACCCACAGAGCACC-3'

[0033] The amplified fragment is 114bp

[0034] GAPDH gene coding region primers:

[0035] SEQ ID NO: 5 Forward: 5'-ATTCCATGGCACCGTCAAGGCTGA-3'

[0036] SEQ ID NO: 6 Reverse: 5'-TTCTCCATGGTGGTGAAGACGCCA-3'

[0037] The amplified fr...

Embodiment 3

[0040] Example 3: Enzyme-linked immunoassay (ELISA) analysis of the expression of TGFβ2 in the supernatants of various breast cancer cell lines

[0041] Prepare the cell culture supernatant: digest the target cells to make a single cell suspension and count them, inoculate 1×106 cells / well in a 6-well plate with a final culture volume of 2ml, and collect the culture supernatant after 48 hours of culture After centrifugation at 2000rpm / min, the supernatant was collected for ELISA experiments.

[0042] Brief steps of ELISA experiment:

[0043] (1) Slowly equilibrate all reagents in the kit to room temperature before use to avoid protein degradation caused by rapid dissolution.

[0044] (2) Preparation of standard substance dilution: add 1ml standard substance dilution solution to the standard substance, let it stand at room temperature for 10min, and rub it repeatedly to promote dissolution. At this time, the concentration is 160ng / ml. First, dilute the standard liquid, and th...

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PUM

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Abstract

The invention belongs to the field of biomedicine and relates to a TGF beta2 gene, a TGF beta 2 polypeptide and application thereof, especially application thereof serving as novel molecular subtyping markers of breast cancer. The circumstance that increase of expression of the TGF beta 2 gene mainly occurs in triple negative breast cancer is verified for the first time, and expression of the TGF beta2 gene in serum of a patient with the triple negative breast cancer is obviously increased than in serum of patients of other molecular subtypes. Novel data are provided for molecular subtyping of the breast cancer, and a novel target and a serologic quick detection method are provided for detecting triple negative molecular subtyping of the breast cancer.

Description

technical field [0001] The present invention belongs to the field of biomedicine, specifically, the present invention relates to TGFβ2 gene, TGFβ2 polypeptide and their applications. Background technique [0002] Breast cancer is one of the malignant tumors with the highest incidence rate and the most serious harm in women. In recent years, with the update of treatment methods, improvement of treatment schemes and diversification of treatment methods, the clinical treatment effect of breast cancer has been greatly improved, but there are still about 500,000 people in the world who die of breast cancer every year. The reason is that breast cancer is a highly heterogeneous disease at the molecular level. The molecular genetic changes of tumors with the same clinical and histopathological morphology are not necessarily similar. These factors affect their treatment and prognosis. Therefore, individualized treatment based on differences in molecular typing is the key to future b...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/12C07K14/495C12Q1/68G01N33/68G01N33/574
Inventor 贺智敏郑国沛张志杰贾小亭
Owner CANCER CENT OF GUANGZHOU MEDICAL UNIV
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