| Lung cancer is one of the main cause leading to death in the cancer-related diseases.The lung cancer morbility and death rate is in the first in male,while second in female.Non-small cell lung cancer(NSCLC)accounts for 80%of all cases of lung cancer.In recent years,it found that long non-coding RNAs(lncRNAs)also play a significant role in the development of tumor.LncRNAs are a class of non-coding RNAs which length is greater than 200 nucleotides.They are important in many life activities such as the dosage compensation effect,epigenetic regulation,cell cycle control,cell differentiation,being a hot topic in the genetic field.It has been specified that tihe lncRNA IRAIn is related to acute myeloid leukemia(AML)and breast cancer in the development of cancer.In our study,we studied IRAIN expression in NSCLC,besides we used experiments in vitro to find it could promote cancer process by affecting cell cycle distribution.Part one IRAIN expression level in NSCLC and the relation to NSCLC clinicopathologic featuresObjective:To determine the expression of IRAIN in NSCLC.Methods:The expression of IRAIN in 47 pairs of tumor tissues of NSCLC and their controls is tested by qRT-PCR.Besids we analyzed the relation between its expression and clinicopathological features(age,gender,smoking history,tumor size,histologial type,tumor stage,tumor grade,lymph node metastasis)of the 47 pairs from NSCLC.Results:1.The expression of IRAIN is overexpressed in tumor tissues of NSCLC compared with their controls.(P<0.05)2.The expression of IRAIN is probably related to tumor size and smoking status,but irrelevant to age,gender,histologial type,tumor stage,tumor grade and lymph node metastasis.(P>0.05)Conclusions:IRIAN may play a role of promoting cancer gene in NSCLC.Part two The effect of IRAIN on cell function in NSCLCObjective:To study the possible role of IRAIN playing in NSCLC devlelopment.Methods:The endogenous IRAIN in NSCLC cell lines was knocked down by chemically synthesized siRNAs,and ectopic overexpressed by transfecting the NSCLC cell lines with the pcDNA3.1-IRAIN expression vector.NSCLC cells transfected Scrambled or Empty vector were as controls.We used cck-8 to determine the proliferation of NSCLC cells,and flow cytometry after staining with annexin V/PI,PI/RNase to analyze cell cycel and cell apoptosis of NSCLC cells transfected with siRNA-IRAIN,pcDNA3.1-IRAIN,Scrambled,and Empty vector.A549 cells were infected by recombinant adenoviral vector producing shRNA and carrying sh-Scrambled as control,which were injected subcutaneously into BALB/C,nu/nu mice to establish A549 cells cancer xenograft models.Tumor volumes were measured once four days since the implantations were starting to grow bigger.Mice from each group were scarified after 32 days of treatment,and the weight of tumor mass was measured.Results:1.CCK-8 assay revealed that the growth was decreased in cells transiently transfected with siRNA compared with its control.At the same time,the overexpression of IRAIN significantly increased the growth of NSCLC cells.2.NSCLC cells transfected with siRNA1 or siRNA2 were arrest in the G0-G1 phase,and the population of cell in the S phase was decreased.In contrast,the population of cell in the S phase was increased in NSCLC cells with overexpressed IRAIN.3.No obvious change of apoptosis ability was observed in A549 and SPCA1 after knockdown by siRNA-IRAIN.4.In vivo experiments showed that the growth of transplanted tumor was restrained after konokdowned by IRAIN ShRNA.Conclusions:These results indicated that IRAIN may promote NSCLC cell into S phase to promote NSCLC cell proliferation. |
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