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Molecular marker for nasopharynx cancer diagnosis and treatment

A nasopharyngeal cancer, one-to-one technology, applied in analytical materials, biological testing, drug combinations, etc., can solve problems such as lack of sensitivity and specificity

Active Publication Date: 2016-02-03
QINGDAO MEDINTELL BIOMEDICAL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although these indicators alone or in combination provide useful information for the diagnosis of NPC, they lack sufficient sensitivity and specificity

Method used

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  • Molecular marker for nasopharynx cancer diagnosis and treatment
  • Molecular marker for nasopharynx cancer diagnosis and treatment
  • Molecular marker for nasopharynx cancer diagnosis and treatment

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0060] Differences in the expression of SMPD3 gene in normal tissue and nasopharyngeal carcinoma tissue in embodiment 1

[0061] 1. Research object:

[0062] 40 cases of patients diagnosed with nasopharyngeal carcinoma and 40 cases of patients diagnosed with chronic inflammation of nasopharyngeal carcinoma (serving as normal nasopharyngeal epithelial tissue control), respectively obtained nasopharyngeal carcinoma tissue and normal nasopharyngeal epithelial tissue by microdissection technology . All clinical samples used in this study were informed to the patients and approved by the Ethics Committee of our hospital.

[0063] 2. Detection of differential expression of SMPD3 gene at the transcriptional level

[0064] 2.1 Extraction of tissue RNA

[0065] 1) In a clean area with less RNase interference, use a mortar containing an appropriate amount of liquid nitrogen to weigh about 20 mg of the isolated tissue sample, and grind it to powder with a pestle;

[0066] 2) Transfer...

Embodiment 2

[0119] Embodiment 2SMPD3 gene expression plasmid construction

[0120] 1. Construction of SMPD3 gene expression vector

[0121] According to the coding sequence of SMPD3 gene (as shown in SEQIDNO.1), design amplification primer, primer sequence is as follows: forward primer is 5'-ATGGTTTTGTACACGAC-3' (SEQIDNO.7), and reverse primer is 5'-CTATGCCTCCTCCTCCCCCGA-3 '(SEQ ID NO. 8). The coding sequence of the full-length SMPD3 gene was amplified from the cDNA library of adult fetal brain (clontech company, catalog number: 638831), and the cDNA sequence was inserted into the restricted endonuclease The obtained recombinant vector pcDNA3.1-SMPD3 was connected to the eukaryotic cell expression vector pcDNA3.1 double-digested with enzymes BamHI and XhoI for subsequent experiments.

[0122] 2. Culture and transfection of nasopharyngeal carcinoma cells

[0123] The human nasopharyngeal carcinoma cell line HONE-1 was cultured in RPMI-1640 medium (purchased from Hyclone) supplemented wi...

Embodiment 4

[0140] Example 4 Effect of SMPD3 gene on the proliferation of nasopharyngeal carcinoma cells

[0141] MTT assay was used to detect the effect of SMPD3 gene on the proliferation ability of nasopharyngeal carcinoma cells.

[0142] 1. Steps: trypsinize each group of cells 12 hours after transfection, make a single cell suspension, inoculate 6000 cells per well in a 96-well culture plate, set 7 time points for each group, and set 6 times for each time point. multiple holes. After the cells adhere to the wall, perform the first test: add 20 μl of 5g / L MTT solution to each well, continue to cultivate for 4 hours, aspirate the medium, add 150 μl of DMSO, carefully pipette to fully dissolve the purple-blue precipitate, and use a microplate reader The absorbance value (A value) was measured at a wavelength of 490 nm. Then every 12h detection 1, continuous measurement 72h, a total of 7 times. This experiment was repeated 3 times.

[0143] 2. Statistical methods

[0144] The experim...

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Abstract

The invention discloses a molecular marker, namely an SMPD3 gene and an expression product thereof which are used for early diagnosis of nasopharynx cancer. QPCR and Western blot methods prove that the expressions of the SMPD3 gene in a nasopharynx cancer patient and a normal person are different, which can be used as an index for early diagnosis of nasopharynx cancer. In addition, the invention further discloses the expression product of the SMPD3 gene, and the expression product can be used as a therapeutic target of nasopharynx cancer for guiding the development of new medicines.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to the application of SMPD3 gene in the diagnosis and treatment of nasopharyngeal carcinoma. Background technique [0002] Nasopharyngeal carcinoma refers to malignant tumors that occur on the roof and side walls of the nasopharyngeal cavity. China is the place with the highest incidence of nasopharyngeal carcinoma in the world, especially in southern China and Guangdong Province, so nasopharyngeal carcinoma is also called "Guangdong tumor" in my country. According to statistics from the Sun Yat-sen University Cancer Center, nearly 3,000 newly diagnosed cases of nasopharyngeal carcinoma are diagnosed each year, and the incidence rate is the first among malignant tumors of the ear, nose and throat. [0003] At present, the treatment effect of early nasopharyngeal carcinoma patients is very good, and the 5-year survival rate can be as high as 90% or more. The curative effect of radiother...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68G01N33/68G01N33/574A61K45/00A61P35/00
CPCA61K45/00C12Q1/6886C12Q2600/158G01N33/57484G01N33/68G01N2333/47
Inventor 杨承刚董东
Owner QINGDAO MEDINTELL BIOMEDICAL CO LTD
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