Expression And Significance Of Two IncRNAs, AHIF And ANRIL, In Gastric Carcinoma

Gastric cancer is the second mortality malignant tumor worldwide, early gastric cancer patients is less than 10%, and the 5-year survival rate is less than 25%. Due to early gastric cancer patients have no obvious symptoms, and most patients do not take the initiative to do gastroscopy, most gastric cancer has entered middle-late state when be diagnosised. Therefore, it becomes a research hotspot to look for reliable biomarkers for early diagnosis in the field of gastric cancer. Long non-coding RNA is a kind of RNA molecules that transcription of this length is more than 200 nucleotides, without encoding protein or encoding very short polypeptide. Studies have shown that lnc RNA regulates gene expression at the epigenetic level, and plays an important role in the occurrence and development of tumor. Recent studies have shown cycle lnc RNA exist stable in serum or plasma, this suggests that the plasma lnc RNA may become a new non-invasive markers in tumor early diagnosis, evaluation of tumor risk and prognosis.Hypoxia inducing factor-1(HIF-1) is the most extensive endogenous transcription factors in a state under hypoxia of mammals and human beings. Antisense hypoxia inducing factor(a HIF) is the opposite transcription of HIF-1, and is able to identify complementary sequence. Studies have found that a HIF’s function of proto-oncogene in liver cancer, kidney cancer, cervical cancer, breast cancer and other malignant tumors.Antisense non-coding RNA in the INK4 locus(ANRIL) is located in chromosome 9q21, and controls cell growth, proliferation, senescence and apoptosis. Recent studies have found that ANRIL can play an important role in tumors like lung cancer, melanoma, liver cancer, leukemia, and so on.Objective:To study the expression of lnc RNA a HIF and ANRIL in gastric cancer tissues, gastric mucosal epithelial cancerization cells, and plasma of gastric cancer patients, explore their contact and analysis the clinical value combined with clinical pathological characteristics.Methods:1、To detect expression levels of a HIF and ANRIL in tissuesCollect gastric cancer tissues and corresponding tissues adjacent to carcinoma of 33 patients with gastric cancer, and use fluorescent quantitative PCR to detect expression levels of lnc RNA a HIF and ANRIL.2、To establish a cell model, and detect expression of a HIF and ANRIL in cellsUsing chemical carcinogen N-methyl-N’-nitro-N-nitrosoguanidine(MNNG) associated with human gastric cancer to induce immortalized gastric mucosal epithelial line GES-1 cancerate, and then get the malignant transformed MC cells. Cells were observed under optical microscope to compare the morphological changes of GES-1 and MC cells, Western blot experiment was used to test the protein expression changes of E-cad, Vimentin and P-STAT3, at the same time fluorescence quantitative PCR was used to detect expression of lnc RNA a HIF and ANRIL in GES-1 and MC cells.3、To detect expression levels of a HIF and ANRIL in peripheral bloodWe collect 51 cases plasma of gastric cancer patients and 51 cases plasma of healthy people watched with gastric cancer patients’ gender and age. Real-time fluorescent quantitative PCR was used to detect expression levels of lnc RNA a HIF and ANRIL in plasma, and analysis a HIF, ANRIL relativity of expression levels in plasma and gastric cancer tissue. Statistics and analysis the relationship between the expression of lnc RNA a HIF, ANRIL and clinicopathological features of gastric cancer patients.Results:1、The result of fluorescent quantitative PCR showed that the expression levels of lnc RNA a HIF and ANRIL in gastric cancer tissues were obviously higher than that of matched tissues adjacent to carcinoma(P both<0.01).2、Optical microscope showed that gastric mucosa epithelial cells GES-1 formed spindly, and grown orderly by single layer; After being dealt with MNNG, MC cells obvious changed in cell morphology. MC cells contacted closely, bigger, out of tentacles, and formed in irregular, multiple antenna, long spindle and karyotype shape.Result of the Western blot experiment showed that gastric cancer associated protein E-cad low expression, Vimentin and P-STAT3 high expression in MC cells. Compared with GES-1 cells, the expression levels of lnc RNA a HIF and ANRIL in MC cells significantly raised(a HIF:P<0.01;ANRIL:P<0.05).3、The expression levels of lnc RNA a HIF and ANRIL in gastric cancer patients plasma significantly increased that of healthy subjects(P both<0.01), and positively related to the expression levels of gastric cancer tissues(a HIF: r=0.721, P<0.01; ANRIL: r=0.719, P<0.01). The expression levels of lnc RNA a HIF and ANRIL in peripheral blood in patients with gastric cancer were not associated with patients’ gender, age, tumor size, TNM staging, infiltration depth and differentiation degree(P>0.05), but related with lymph node metastasis(P<0.05).Conclusion:In gastric cancer tissues, gastric epithelial cancerization cells and gastric cancer patients’ plasma, lnc RNA a HIF ANRIL expressed highly, and expression level in plasma and in gastric cancer tissues were positively correlated; the expression level of a HIF and ANRIL in peripheral blood in patients with gastric cancer is not related with patients’ gender, age, tumor size, TNM staging, infiltration depth and differentiation degree, but associated with lymph node metastasis. We may provide new clues for clinical diagnosis of gastric cancer through detecting peripheral blood lnc RNA a HIF and ANRIL expression.