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Non-small-cell lung carcinoma molecular marker and applications thereof

A non-small cell lung cancer and molecular marker technology, applied in the field of biomedicine, can solve the problems of high missed detection rate in early censuses, difficulty in finding tumors, and ineffective diagnosis of lung cancer

Pending Publication Date: 2018-07-06
INST OF BASIC MEDICINE OF SAMS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, these methods have their own shortcomings, such as imaging techniques are difficult to find small tumors, and the missed detection rate of early general screening is high
Although the detection effect of lung cancer markers is better than that of imaging techniques, the existing lung cancer markers are broad-spectrum and cannot effectively diagnose lung cancer. Therefore, it is of great significance to find specific lung cancer biomarkers

Method used

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  • Non-small-cell lung carcinoma molecular marker and applications thereof
  • Non-small-cell lung carcinoma molecular marker and applications thereof
  • Non-small-cell lung carcinoma molecular marker and applications thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Example 1 Expression of MAL2 gene and protein in non-small cell lung cancer tissue

[0036] Total RNA was extracted from lung tissue and cDNA was synthesized using Superscript III Kit (Invitrogen), and the prepared cDNA was used as a template for quantitative PCR reaction. Human glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was used as an internal reference, and its upstream and downstream primers were 5'-GCGACACCCACTCCTCCCACCTTT-3';

[0037] 5'-TGCTGTAGCCAAATTCGTTGTCATA-3'. The reaction program is 5 minutes at 95°C; 15 seconds at 95°C, 30 seconds at 58°C, and 30 seconds at 68°C, 40 cycles. The instrument used was ABI 7500 quantitative PCR instrument. The transcription level of MAL2 gene in 28 pairs of non-small cell lung cancer tissues was detected by fluorescent quantitative PCR.

[0038] Depend on figure 1 (A)-(C) It can be seen that the transcription level of MAL2 gene in 28 pairs of non-small cell lung cancer tissues was detected by fluorescent quantitative ...

Embodiment 2

[0039] Example 2 MAL2 promotes lung cancer cell proliferation test

[0040] The plasmid DNA containing the MAL2 gene was transferred into CRL-5872 and A549 lung cancer cells by conventional method liposome transfection method, and the protein expression of the MAL2 gene was verified by Western blot method after transfer into CRL-5872 and A549 cells. Cell proliferation was detected by MTS method.

[0041] figure 2 Western blot in A demonstrates overexpression of exogenously transferred MAL2 in CRL-5872 lung cancer cells; by figure 2 B It can be seen that MAL2 promotes the proliferation of CRL-5872 lung cancer cells after overexpression by detecting cell growth for 6 consecutive days; figure 2 In C, western blotting was used to demonstrate the overexpression of exogenously transferred MAL2 in A549 lung cancer cells; figure 2 In D, by continuously detecting the growth of cells for 6 days, it can be seen that MAL2 promotes the proliferation of A549 lung cancer cells after o...

Embodiment 3

[0042] Example 3 MAL2 transcription inhibition test

[0043] Cultivate A549 cells, according to the full-length sequence of human MAL2 mRNA (NM_052886) published by GenBank, follow the conventional design principles of interfering RNA, design three interfering RNAs namely shMAL2-1, shMAL2-2, shMAL2-3 and design empty vector (EV) according to conventional methods Transfection was carried out, and the interference efficiency of shMAL2-1, shMAL2-2, and shMAL2-3 was detected by fluorescent quantitative PCR technology, and the cell proliferation was detected by MTS method.

[0044] image 3 A Quantitative PCR verified the effective inhibitory effect of three shRNAs of the MAL2 gene on the transcription of MAL2, wherein shMAL2-1, shMAL2-2, and shMAL2-3 represent three different interfering RNAs, and EV represents an empty vector. image 3 A It can be seen that shMAL2-1, shMAL2-2, shMAL2-3 can effectively inhibit the transcription of MAL2; image 3 B shows that through continuous d...

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Abstract

The invention belongs to the field of biomedicine, and relates to a non-small-cell lung carcinoma molecular marker and applications thereof. According to the present invention, the deep research results show that T cell differentiation protein 2 (MAL2) gene is closely related to the occurrence of non-small-cell lung carcinoma, such that the T cell differentiation protein 2 (MAL2) gene can be usedas the molecular marker for early diagnosis of non-small-cell lung carcinoma; by determining the functions of the key pathogenic gene in non-small-cell lung carcinoma, the molecular target in non-small-cell lung carcinoma can be determined so as to accelerate the progress of the individualized targeted therapy of non-small-cell lung carcinoma; specifically the present invention discloses applications of the MAL2 gene and protein in gene molecular markers for the detection of non-small-cell lung carcinoma, and provides applications of the MAL2 gene and protein in preparation of clinical diagnostic reagents or kits for tumors; and the new gene and the protein highly expressed in non-small-cell lung carcinoma are found so as to provide the new approach for the diagnosis and treatment of tumors.

Description

technical field [0001] The invention belongs to the field of biomedicine, and relates to a non-small cell lung cancer molecular marker and application thereof. Background technique [0002] The morbidity and mortality of lung cancer are very high worldwide. According to the statistical data of 2012, the International Agency for Research on Cancer estimated that lung cancer was the cancer with the highest mortality rate in that year. About one in five cancer deaths died from lung cancer (accounting for 19.4% of fatalities). [0003] Lung cancer can be roughly divided into two categories based on histopathological differences: small cell lung cancer and non-small cell lung cancer. About 85% of lung cancers are non-small cell lung cancers. It is further subdivided into three categories: adenocarcinoma, squamous cell carcinoma, and large cell carcinoma, which are driven by multiple activated oncogenes. The five-year survival rate for patients with non-small cell lung cancer i...

Claims

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Application Information

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IPC IPC(8): C12Q1/6886G01N33/68G01N33/574
CPCC12Q1/6886C12Q2600/136C12Q2600/158G01N33/57423G01N33/6884G01N2333/47G01N2500/00
Inventor 刘红艳姜国胜崔大勇姚成芳韩杨李娟
Owner INST OF BASIC MEDICINE OF SAMS
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