阅读说明：本技术 lncRNA IGFL2-AS1作为结肠癌诊断标志物的应用 (Application of lncRNA IGF L2-AS 1 AS colon cancer diagnosis marker ) 是由 李小俊 毕焕京 闫飞 李倩 于 2020-03-20 设计创作，主要内容包括：本发明公开了一种lncRNA IGFL2-AS1作为结肠癌诊断标志物的应用,属于肿瘤分子生物学领域。本发明提供的lncRNA IGFL2-AS1在结肠癌中呈现高水平表达,在结肠癌细胞恶性转化及肿瘤形成过程中发挥重要作用。人为敲低结肠癌细胞中的lncRNA IGFL2-AS1,能够明显抑制SW620细胞的增殖、迁移、侵袭能力和恶性程度,表明lncRNA IGFL2-AS1在结肠癌发病过程中发挥重要作用,可以作为诊疗结肠癌的新的分子标记和药物靶点。(The invention discloses an application of lncRNA IGF L-AS 1 AS a colon cancer diagnosis marker, belonging to the field of tumor molecular biology.A lncRNA IGF L2-AS 1 provided by the invention shows high-level expression in colon cancer and plays an important role in the processes of malignant transformation of colon cancer cells and formation of tumors.)

The application of lncRNA IGF L2-AS 1 AS a colon cancer diagnosis marker.

2. The use of claim 1, wherein the incrna IGF L2-AS 1 has the nucleotide sequence shown in SEQ ID No. 1.

3. Application of a reagent for detecting lncRNA IGF L2-AS 1 in preparation of a colon cancer diagnosis detection reagent or kit.

4. The use of claim 3, wherein the detection reagent or kit is used for detecting the expression level of IncRNA IGF L2-AS 1 in a biological sample.

5. The use according to claim 4, wherein the detection reagent or kit comprises PCR primers with detection specificity for IncRNAIGF L2-AS 1.

6. The use of claim 5, wherein the PCR primers with detection specificity for IncRNA IGF L2-AS 1 are AS follows:

an upstream primer: 5'-TGCTGAGCAC AGACATGTGA-3', respectively;

a downstream primer: 5'-GTGCTGCAGA ATCAACGACC-3' are provided.

7. A group of interfering RNAs aiming at lncRNA IGF L2-AS 1 gene targets is characterized in that the sequences of the lncRNA IGF L2-AS 1 gene targets are shown in SEQ ID NO. 1, and the interfering RNAs comprise at least one of siRNA-1, siRNA-2 and siRNA-3.

8. The interfering RNA of claim 7, wherein the sequence of siRNA-1 is: 5'-AAAAUUGGUGAGAAGUUCCUU-3' are provided.

9. The interfering RNA of claim 7, wherein the sequence of siRNA-2 is: 5'-UAAAAUUGGUGAGAAGUUCCU-3' are provided.

10. The interfering RNA of claim 7, wherein the sequence of siRNA-3 is: 5'-ACUUCUUCAUUCUUGUUGGCU-3' are provided.

Technical Field

The invention belongs to the field of tumor molecular biology, and particularly relates to application of lncRNA IGF L2-AS 1 AS a colon cancer diagnosis marker.

Background

Worldwide, the incidence of colorectal cancer is third in the malignancy, and mortality is second, and is considered a major threat to public health. Due to poor eating patterns and living habits, the incidence and mortality of colon cancer presents a significantly rising trend and is one of the most common gastrointestinal malignancies. In China, about 17 ten thousand new cases of colon cancer occur each year, and the patients are younger in disease onset age, which causes great harm to the health of people in China. Colorectal carcinogenesis is a multistep biological process involving the dysregulation of multiple oncogenes and tumor suppressor genes. Despite the relatively deep understanding of the disease colorectal cancer, the continuous progress of diagnostic techniques and the increasing efficiency of therapeutic measures, the overall survival rate of colorectal cancer patients remains relatively low. At present, with the increasing incidence and poor prognosis of colorectal cancer, there is an urgent need to understand the specific molecular mechanisms involved in the pathogenesis of colorectal cancer more deeply, since this may facilitate the development of more effective therapeutic strategies, ultimately improving the prognosis of the patient.

To date, with improvements in whole genome and transcriptome sequencing technologies, it has been found that the whole genome of most mammals can be transcribed, but most of the transcripts have limited or no protein-encoding capacity at all, and these genes are referred to as ncrnas. IncRNAs are non-coding RNAs of length greater than 200 nucleotides that are transcribed by RNA polymerase II (RNAPII), and which have few or no Open Reading Frames (ORFs). A great deal of research results show that lncRNA plays an irreplaceable role in various basic biological processes such as embryonic development, epigenetic silencing, transcription and translation control, various biological behaviors of cells, tumorigenesis and the like. In addition, extensive research has provided strong evidence that lncRNA plays a functional role in a range of human diseases, including various types of cancer, such as breast, pancreatic, lung, gastric, and the like. And a plurality of lncRNA are also found to participate in carcinogenesis and cancer suppression processes in the process of generating and developing tumors, for example, lncRNA MVIH can be used AS an oncogene to promote the proliferation and invasion of non-small cell lung cancer cells, and on the contrary, lncRNA KCNK15-AS1 is proved to play a role in protecting the non-small cell lung cancer cells AS a cancer suppressor. Recent studies have shown that lncrnas also play a role as modulators in the development and progression of colorectal cancer, suggesting their potential as new therapeutic targets. Research on the molecular mechanisms and biological functions of lncRNA in the development and progression of colorectal cancer is still in the stage of inception, and although researchers have had limited success to date in elucidating specific molecular mechanisms, these limited studies are sufficient to demonstrate that lncRNA is a potential biological target for colorectal cancer diagnosis and treatment.

Therefore, the invention provides a breakthrough point for treating the colon cancer by detecting the expression and the function of lncRNA IGF L2-AS 1 in the colon cancer.

Disclosure of Invention

The invention aims to provide application of lncRNA IGF L2-AS 1 AS a colon cancer diagnosis marker.

The technical scheme adopted by the invention is as follows:

application of lncRNA IGF L2-AS 1 AS colon cancer diagnosis marker.

Furthermore, the nucleotide sequence of lncRNA IGF L2-AS 1 is shown in SEQ ID NO. 1.

Application of a reagent for detecting lncRNA IGF L2-AS 1 in preparation of a colon cancer diagnosis detection reagent or kit.

Further, the detection reagent or the kit is used for detecting the expression level of lncRNA IGF L2-AS 1 in the biological sample.

Further, the detection reagent or the kit comprises a PCR primer with detection specificity to lncRNA IGF L2-AS 1.

Further, PCR primers with detection specificity for lncRNA IGF L2-AS 1 were AS follows:

an upstream primer: 5'-TGCTGAGCAC AGACATGTGA-3', respectively;

a downstream primer: 5'-GTGCTGCAGA ATCAACGACC-3' are provided.

The sequences of lncRNA IGF L-AS 1 gene targets are shown in SEQ ID NO: 1, and the interfering RNAs comprise at least one of siRNA-1, siRNA-2 and siRNA-3.

Further, the sequence of siRNA-1 is: 5'-AAAAUUGGUGAGAAGUUCCUU-3' are provided.

Further, the sequence of siRNA-2 is: 5'-UAAAAUUGGUGAGAAGUUCCU-3' are provided.

Further, the sequence of siRNA-3 is: 5'-ACUUCUUCAUUCUUGUUGGCU-3' are provided.

Compared with the prior art, the invention has the beneficial effects that:

artificially knocking down lncRNA IGF L2-AS 1 in colon cancer cells can obviously inhibit the proliferation, migration, invasion capacity and malignancy of SW620 cells, shows that lncRNA IGF L2-AS 1 plays an important role in the colon cancer morbidity process, and can be used AS a new molecular marker and a drug target for diagnosing and treating colon cancer.

Drawings

The accompanying drawings, which are included to provide a further understanding of the invention and are incorporated in and constitute a part of this specification, illustrate embodiments of the invention and together with the description serve to explain the invention and not to limit the invention. In the drawings:

FIG. 1 is a Real-time PCR assay of the expression of IncRNA IGF L2-AS 1 in colon cancer and paracarcinoma tissues;

FIG. 2 shows the expression of IncRNA IGF L2-AS 1 in SW620 cell line with knockdown of IncRNA IGF L2-AS 1;

FIG. 3 shows the CCK-8 proliferation of SW620 cell line knocking down IncRNA IGF L2-AS 1;

FIG. 4 shows the results of cell migration of SW620 cell line with knockdown of IncRNA IGF L2-AS 1;

FIG. 5 shows the results of cell invasion experiments on SW620 cell line with knockdown of IncRNA IGF L2-AS 1.

Detailed Description

The invention utilizes fluorescent quantitative PCR experiment to research that the expression quantity of lncRNA IGF L2-AS 1 in colon cancer is obviously higher than that of tissues beside cancer, extracts total RNA of the colon cancer and tissues beside cancer, carries out reverse transcription to synthesize cDNA, designs lncRNA IGF L2-AS 1PCR primer to carry out PCR amplification, and clones for the first time to obtain lncRNA IGF L2-AS 1 gene.

By designing siRNA, SW620 colon cancer cell line with low lncRNA IGF L2-AS 1 is obtained, and the influence of lncRNA IGF L2-AS 1 on the proliferation, migration, invasion and malignancy of SW620 cells is researched, so that the effect of lncRNA IGF L2-AS 1 in the colon cancer pathogenesis process is discussed, and the application of the lncRNA IGF L-AS 1 AS a new tumor marker in colon cancer clinical diagnosis, treatment and drug development is discussed.

The embodiments of the present invention are described below with reference to specific embodiments, and other advantages and effects of the present invention will be easily understood by those skilled in the art from the disclosure of the present specification. The invention is capable of other and different embodiments and of being practiced or of being carried out in various ways, and its several details are capable of modification in various respects, all without departing from the spirit and scope of the present invention.

In the following embodiments, unless otherwise specified, the technical means used are conventional means well known to those skilled in the art.