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MicroRNA marker for identifying subtypes of lung cancer and application of microRNA marker

A technology for lung cancer and purpose, applied in the field of diagnosis of lung cancer subtypes, to achieve the effects of low-cost targeted therapy, broad application prospects, and accurate and reliable results

Inactive Publication Date: 2012-12-26
ZHONGSHAN HOSPITAL FUDAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are few reports on microRNAs that can accurately diagnose lung cancer subtypes obtained from pure target cells isolated from lung cancer tissues.

Method used

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  • MicroRNA marker for identifying subtypes of lung cancer and application of microRNA marker
  • MicroRNA marker for identifying subtypes of lung cancer and application of microRNA marker
  • MicroRNA marker for identifying subtypes of lung cancer and application of microRNA marker

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Example 1 Screening acquisition and independent verification of microRNA

[0043] The overall experimental plan is as follows:

[0044] By collecting 586 lung cancer tissue or exfoliated cell samples from Zhongshan Hospital between August 2008 and December 2011, each case was reviewed by two experienced lung cancer pathologists, and 586 samples were assigned to one of five groups, among which The cases in each group were independent and did not overlap.

[0045] 1. Discovery group: using laser capture microseparation technology to extract cancer cells from 82 cases (36 cases of AC, 30 cases of SQ and 16 cases of SCLC) from independently frozen tissues for microarray research, and obtained candidate microRNAs for lung cancer typing, And further confirmed its value by quantitative reverse transcription-polymerase chain reaction (RT-PCR).

[0046] 2. Training group: 85 paraffin-embedded lung tissues (29 AC, 24 SQ, and 32 SCLC) were manually isolated to ensure > 75% can...

Embodiment 2

[0120] Example 2 Clinical application of microRNA combination A and combination B

[0121] Two cytopathologists from Shanghai Cancer Hospital and Zhongshan Hospital carried out the lung cancer typing of bronchial brushing cell samples, and directly compared the diagnostic accuracy of cell typing and microRNA combination typing. The results are shown in Table 5. It can be seen from the table that although microRNA combination A (has-miR-29a and has-miR-375) failed to improve the diagnosis of NSCLC compared with cytological diagnosis, it could promote the diagnosis of SCLC; at the same time, the combination of microRNA B (has-miR-205 and has-miR-34a) could more effectively discriminate AC from SQ. The combination of cytology and microRNA detection can greatly improve the diagnosis of lung cancer subtypes, reduce the number of undiagnosed patients by cytology, and help solve the problem of inability to accurately diagnose lung cancer subtypes by brushing cytology. The combinat...

Embodiment 3

[0125] Example 3 Preparation of microRNA chip

[0126] Convert the microRNA (SEQ ID No.2, 3, 4, 7) provided in Table 1 into a complementary sequence, and add a 10-20nt connection sequence at both ends of the sequence according to the GC alignment characteristics of the generated sequence; the core sequence is different, and the connection sequence The sequence is also different. The junction sequence can be randomly generated by the program, and the probes formed by the junction sequence and the core sequence meet the following conditions:

[0127] 1) In the probe sequence, the number of the same nucleotide (A, C, G, T) cannot exceed 50% of the total number of sequences;

[0128] 2) The number of any consecutive A, T or C, G cannot exceed 50% of the total number of sequences;

[0129] 3) G and C content account for 40%-60% of the total number of sequences;

[0130] 4) The probe sequence cannot self-hybridize, that is, the length of the complementary fragment in the probe ...

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Abstract

The invention provides microRNA (micro ribonucleic acid) for identification of subtypes of a lung cancer. The miroRNA is chosen from the following sequence: a) a sequence shown as SEQ ID No. n (sequence identification number), or b) microRNA complementary to the sequence shown as SEQ ID No. n, wherein the n is 2, 3, 4 or 7. The invention further provides an application of the microRNA and a reagent kit based on the microRNA. The invention further provides a set formed by the microRNA, an application of the set and a reagent kit based on the set. The miroRNA has the advantages that the microRNA can form two combinations, namely SEQ ID No.2 and SEQ ID No.7, as well as SEQ ID No.3 and SEQ ID No.4, which can be used to differentially diagnose the small cell lung cancer, non-small cell lung cancer, adenocarcinoma lung cancer and squamous cell lung cancer respectively; the diagnosis is high in speed, low in cost and accurate and reliable in result; and a significant basis is provided for performing precise targeted therapy on a patient with the lung cancer.

Description

technical field [0001] The present invention relates to the diagnosis of lung cancer subtypes, in particular to a microRNA marker for distinguishing lung cancer subtypes and the application of the microRNA. Background technique [0002] Lung cancer has become the malignant tumor with the highest mortality rate in the world. The latest data from the World Health Organization in 2011 shows that in 2008, there were about 1.6 million new lung cancer patients and about 1.4 million lung cancer patients died. Among lung cancer patients, small cell lung cancer (SCLC) accounts for about 15%, and non-small cell lung cancer (NSCLC) accounts for about 80%. NSCLC mainly includes adenocarcinoma lung cancer (adenocarcinoma lung cancer, AC) and squamous cell lung cancer (SQ) two types. Lung cancer is one of the malignant tumors with the worst prognosis. Even if early-stage (pIa-pIb) lung cancer is treated with surgery, radiotherapy and chemotherapy, the 5-year recurrence rate is still as h...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/113C12Q1/68C40B40/06
Inventor 卢韶华白春学黄威侯英勇吴莹朱虹光
Owner ZHONGSHAN HOSPITAL FUDAN UNIV
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