| Objective: Gastric cancer is one of the most frequently occurring cancers in China.To investigate its possible biomarker can deepen the understanding of the pathogenesis of gastric cancer and reasonably evaluate its potential and value in the diagnosis and treatment of gastric cancer.Long-chain non-coding RNA(lnc RNA)has been demonstrated to play an important role in the development of many diseases,including the occurrence of malignant tumors,and it can also be used as a molecular marker for early diagnosis of gastric cancer.In this study,the unexplored target lnc RNA was selected from the chip test results obtained in the early stage of the experiment from our group for further research,its expression level was verified by expanding the sample size of gastric adenocarcinoma tissues,and its function in gastric adenocarcinoma cells was preliminarily explored.Methods: According to the previous lnc RNA chip test results of the experiment from our group,selecting the target lnc RNA RP11-325L7.1 for further study.(1)Using reverse transcription real-time fluorescent quantitative PCR(RT-q PCR)to detect lnc RNA RP11-325L7.1 expression levels in 119 pairs of gastric adenocarcinoma tissues and adjacent non-cancerous tissues.The expression levels of lnc RNA RP11-325L7.1 were then compared with the chip results;the basic pathological information of patients and the results of immunohistochemical markers were collected,and the correlation between lnc RNA RP11-325L7.1 expression levels and them was analyzed statistically.(2)Constructing the lnc RNA RP11-325L7.1overexpression plasmid vector.The plasmid was then transfected into cells and screened for G418 resistanceso that gastric cancer cells which were stably overexpressed lnc RNA RP11-325L7.1 were obtained.Cell biological behaviors such as cell proliferation,migration,invasion,and apoptosis after up-regulation of RP11-325L7.1 expression were analyzed by CCK-8 experiment,plate cloning experiment,Transwell migration and invasion experiment and flow cytometry.(3)Through ce RNA network and CNC analysis,the correlation between target lnc RNA RP11-325L7.1 and its action targets on lnc RNA-mi RNA-m RNA net,pathway and lnc RNA-m RNA were analyzed.Results:(1)The expression profile chip results showed that the target lnc RNA RP11-325L7.1 was significantly down-regulated in 6 gastric adenocarcinoma tissues.The q RT-PCR results showed that when compared with that in the paired non-cancerous tissues,lnc RNA RP11-325L7.1 was down-regulated in 89 of 119 cases of gastric adenocarcinoma tissues,and the down-regulated expression rate was 74.79%(89/119)with an average down-regulation folds of 3.34 times.The difference was statistically significant(P<0.01).The q RT-PCR verification results exhibited the same trend change compared with that detected by the chip.(2)The expression levels of lnc RNA RP11-325L7.1 in gastric adenocarcinoma tissues were correlated with the lymph node metastasis of patients with this type of cancer(P=0.028).(3)Constructing lnc RNA RP11-325L7.1 overexpression plasmid vector and selecting gastric cancer cells MGC-803 and AGS cell clones with stably overexpressing lnc RNA RP11-325L7.1 for cell function experiments in vitro.The results showed that increasing the expression of lnc RNA RP11-325L7.1 inhibited both MGC-803 and AGS cell proliferation,colony formation,migration and invasion(P<0.01).Overexpression of lnc RNA RP11-325L7.1 could promote AGS cell apoptosis(P<0.05).Conclusions: LncRNA RP11-325L7.1 is down-regulated in gastric adenocarcinoma tissues and cells;overexpression of lnc RNA RP11-325L7.1 leads to inhibit the proliferation,clone formation,migration and invasion of MGC-803 and AGS gastric adenocarcinoma cells and promote AGS cell apoptosis.Therefore,lnc RNA RP11-325L7.1 may be used as a potential molecular marker for gastric cancer research. |
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