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A secreted sema4c protein and its application

A secreted, protein-based technology, applied in the field of medical biology, can solve problems such as elevated levels, significant differences in expression, and inconvenient use, and achieve the effects of less pain, low cost, and good specificity

Active Publication Date: 2017-11-17
WUHAN KDWS BIOLOGICAL TECH CO LTD
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0010] In view of the problems in the prior art, the present invention provides a secreted Sema4C protein, which not only confirms the existence of the secreted Sema4C protein for the first time, but also finds that the expression of the secreted Sema4C protein in the serum of normal people and tumor patients is significantly different , especially in the serum of patients with lymph node metastasis of breast cancer, and it is significantly positively correlated with the tumor burden of breast cancer patients, so it can be used as an ideal serological marker for preoperative diagnosis and postoperative treatment of breast cancer patients Effect evaluation and follow-up monitoring, further providing a kit using secreted Sema4C protein as a tumor marker, which solves the problem that membrane-type Sema4C protein cannot be detected by Elisa and must use immunohistochemical technology.

Method used

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  • A secreted sema4c protein and its application
  • A secreted sema4c protein and its application
  • A secreted sema4c protein and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0061] The level of SEMA4C nucleic acid in tumor tissue was significantly up-regulated, and the expression of membrane-type Sema4C protein in breast cancer tissue was up-regulated.

[0062] The previous inventors used LCM technology to capture homogeneous lymphatic endothelial cells from the frozen tissue sections of breast cancer primary tumors and normal breasts, and the micro-extracted RNA was unbiased and linearly amplified to obtain sufficient and complete RNA , effectively coupled with Affymetrix GeneChip array HG U133Plus 2.0 polyoligonucleotide expression profiling chip, to screen functional gene groups differentially expressed in lymphatic endothelial cells between breast cancer tissue and normal breast tissue (Wu, M., Han, L., Shi ,Y.,Xu,G.,Wei,J.,You,L.,Chen,Y.,Zhu,T.,Li,Q.,Li,S.,et al.(2010).Development and characterization of a novel method for the analysis of gene expression patterns in lymphatic endothelial cells derived from primary breast tissues. J Cancer Res...

Embodiment 2

[0067] Example 2: Analysis of the relationship between membrane-type Sema4C expression and clinicopathological parameters

[0068]The relationship between the expression of membrane-type Sema4C in breast cancer tissues and clinicopathological features is shown in Table 2. Chi-square test analysis showed that the positive expression of Sema4C in the model was more likely to occur in patients with lymph node metastasis and breast cancer tissue type Ⅲ. The incidence of lymph node metastasis in the membrane-type Sema4C positive expression group was 82.9% (68 / 82), which was significantly higher than that in the non-metastatic group (13.6% (11 / 81). In addition, the distribution rate of tissue types was also different between the two groups. The positive rate of Sema4C expression in patients with breast cancer tissue type Ⅰ and breast cancer tissue type Ⅱ was 47.9% (59 / 123) and breast cancer tissue type Ⅱ. The positive rate of expression in patients with tissue type III was 77.5% (3...

Embodiment 3

[0071] Example 3: Discovery of the presence of secreted Sema4c protein

[0072] step 1

[0073] Digest and collect MD-MBA-231 cells (from ATCC) with EDTA-containing trypsin (purchased from Gibco, trade name TYPSIN, wherein the mass volume ratio of trypsin is 0.25%), and the adherent cells are removed from the culture flask The wall fell off. Wash with 1×PBS (the composition is 140mM NaCl, 2.7mM KCl, 10mM NaCl 2 HPO 4 , 1.8mM KH 2 PO 4 , pH value is 7.4), centrifuge at 1000rpm for 5 minutes, discard the supernatant, add protein lysate (Biyuntian P0013) according to the amount of cells, lyse for 30min, 12,000rpm at 4°C, centrifuge for 15 minutes, take the supernatant and stain with Coomassie Brilliant Blue OD was measured by UV spectrophotometry 595 After the value, the concentration was calculated according to the standard curve, and the protein quantification was 30ug and 50ug, respectively, and then placed at -80°C for use to obtain the total protein of the cells.

[0...

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Abstract

The invention relates to secreting type Sema 4C protein and application thereof. The secreting type Sema 4C protein is an extracellular section of membrane type Sema 4C protein and comprises amino acid sequences shown in the SEQ ID NO.1. The secreting type Sema 4C protein can be used as a tumor marker to be applied. The invention further provides a reagent kit adopting the secreting type Sema 4C protein as the tumor marker and adopting Sema 4C human recombinant protein as the standard substance, and the amino acid sequence is shown in SEQ ID NO.2. The reagent kit adopting the secreting type Sema 4C protein as the tumor marker can be used for preoperative diagnosis, aftertreatment effect evaluation and follow-up visit monitoring of a cancer patient (particularly a breast cancer patient).

Description

technical field [0001] The present invention belongs to the field of medical biotechnology, and in particular relates to a newly discovered secreted Sema4C protein and its application, specifically an ELISA using secreted Sema4C protein as a tumor marker that can be used for early detection and postoperative monitoring of breast cancer (Enzyme-linked immunosorbent assay) kit preparation and application. Background technique [0002] The incidence of breast cancer ranks first in female patients, accounting for about 23% of all malignant tumors suffered by women. According to the data from the National Health and Family Planning Commission, from 2004 to 2005, the mortality rate of breast cancer in Chinese women was 5.9 / 100,000, ranking the 6th in the mortality rate of malignant tumors. In the past 15 years, the age-adjusted rate of incidence and death of breast cancer and the absolute number of deaths have increased significantly in my country. Faced with such a severe situa...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/435
CPCG01N33/57415
Inventor 马丁高庆蕾魏军成杨洁乔龙
Owner WUHAN KDWS BIOLOGICAL TECH CO LTD
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