Expression Of Long Non-Coding RNA FAM83H-AS1 In Gastric Cardia Adenocarcinoma And Its Biological Effects

Objective: To investigat the expression of long non-coding RNA(lncRNA)FAM83H-AS1 in gastric cardia adenocarcinoma(GCA)and gastric cancer cell lines,and to analyze the correlation between its expression and clinicopathological parameters.To study the effects of knockdown of lncRNA FAM83H-AS1 on proliferation,migration and invasion of gastric cancer cell lines in vitro.Methods:1.Real-time quantitative polymerase chain reaction(RT-qPCR)method was used to detect the expression of lncRNA FAM83H-AS1 in 60 cases of GCA tissues and corresponding adjacent normal tissues,and in four human gastric cancer cell lines(BGC-823,SGC-7901,MGC-803,HGC-27).2.Design and synthesize a shRNA that specifically knockdown FAM83H-AS1,and transfected to gastric cancer cell lines BGC-823 and SGC-7901 that knockdown the lncRNA FAM83H-AS1 by lipofection.Transfection efficiency was measured by RT-qPCR method.3.The effect of knockdown of lncRNA FAM83H-AS1 on proliferation of gastric cancer cells was detected by MTS and clone formation methods.4.Using the scratch test to detect the effect of knocking down lncRNA FAM83H-AS1 on the migration ability of gastric cancer cells.5.Transwell chamber invasion assay was used to detect the invasive ability of gastric cancer cells after knockdown the lncRNA FAM83H-AS1.Result:1.Expression of lncRNA FAM83H-AS1 in GCA tissues and corresponding adjacent normal tissues: the expression of lncRNA FAM83H-AS1 in GCA tissues was significantly higher than that in corresponding normal tissues.The expression was closely related to TNM stage,lymph node metastasis and degree of differentiation(P<0.05),but not related to age and gender(P>0.05).2.Expression of lncRNA FAM83H-AS1 in gastric cancer cell lines: lncRNA FAM83H-AS1 was highly expressed in other cell lines except for the lowest expression in HGC-27 cell line,among which BGC-823 cell line was the highest.3.Screen and verify knockdown efficiency of lncRNA FAM83H-AS1 knockdown plasmid: lncRNA FAM83H-AS1 knockdown of BGC-823 and SGC-7901 cells were successfully constructed,and the knock down efficiency was detected by RT-qPCR.The relative expression of lncRNA FAM83H-AS1 in sh1 plasmid transfection group was the lowest,knockdown efficiency was the highest(P<0.05).Therefore,sh1 plasmid was selected for subsequent experiments.4.The effect of knocking down FAM83H-AS1 on proliferation of gastric cancer cells: MTS and clone formation assays showed that knockdown of lncRNA FAM83H-AS1 inhibited BGC-823 and SGC-7901 cells proliferation ability(P<0.05).5.The effect of knockdown FAM83H-AS1 on migration ability of gastric cancer cells: Knockdown of lncRNA FAM83H-AS1 had no inhibitory effect on the migration ability of BGC-823 cells(P>0.05),which inhibited SGC-7901 cell migration ability(P<0.05).6.The effect of knocking down lncRNA FAM83H-AS1 on the invasion ability of gastric cancer cells: Knockdown of lncRNA FAM83H-AS1 inhibited the invasive ability of BGC-823 and SGC-7901 cells(P<0.05).In conclusion:1.The long non-coding RNA FAM83H-AS1 is highly expressed in gastric cardia adenocarcinoma tissues and gastric cancer cell lines,which may be related to the occurrence and development of gastric cardia adenocarcinoma.2.Long non-coding RNA FAM83H-AS1 can affect the proliferation,migration and invasion of gastric cancer cells in vitro.