Effects Of PDIA6 On Proliferation And Migration Of Oral Squamous Cell Carcinoma Cells Through MAPK/JNK

Objective: To investigate the effect of PDIA6 gene on the proliferation and migration of OSCC cells and their possible molecular mechanisms,in order to provide a theoretical basis for targeted therapy.Methods: After silencing the PDIA6 gene of human squamous cell carcinoma of tongue cells CAL27 and TCA8113 by lipofection,qRT-PCR method and the Western Blot were used to detect the degree of gene silencing.After the PDIA6 silencing,we used MTT method to detect the ability of cell proliferation,and we used Wound healing assay to measure the migration ability of cells.Finally,qRT-PCR method was used to detect the m RNA expression levels of MAPK and JNK genes after PDIA6 silencing,and Western Blot was used to detect the expression of MAPK,JNK and pho-JNK proteins.Results:1.(1)The qRT-PCR resulted: After transfection of PDIA6-si RNA in CAL27 and TCA8113 cells for 24 hours,compared with the negative control group,the relative m RNA expression levels of PDIA6 gene were significantly down-regulated in the two cells after transfection for 24 hours,and the difference was statistically significant(P<0.01).(2)Western blotting experiment resulted: The relative protein expression level of PDIA6 gene was significantly lower compared with the negative control group,and the difference was statistically significant(P<0.01).2.MTT experiment resulted: After transfection of PDIA6-si RNA in CAL27 and TCA8113 cells for 24 hours,the culture was continued for 48 and 72 hours.Compared with the negative control group,the proliferation rate of the two cells were significantly reduced at 24 hours,48 hours and 72 hours.With the prolongation of time,the proliferation inhibitory effect was more obvious,and the differences above were statistically significant(P <0.05).3.Wound healing assay resulted: The CAL27 and TCA8113 cells were cultured for 24 and 48 hours after they stably transfected PDIA6-si RNA for 24 hours.The wound healing area of the transfection group significantly decreased compared with the negative control group,and the difference was statistically significant(P<0.05).4.The qRT-PCR experiment resulted: 24 hours after the CAL27 and TCA8113 cells were stably transfected PDIA6-si RNA,compared with the negative control group,the relative m RNA expression levels of MAPK and JNK were significantly down-regulated.Western blotting experimental resulted: 24 hours after PDIA6-si RNA transfection of two kinds of cells,the relative proteins expression levels of MAPK and JNK were significantly down-regulated compared with the negative control group,the relative expression level of phosphorylated JNK protein was also significantly down-regulated,and the differences above were statistically significant(P<0.05).And it further proved that the MAPK/JNK was inhibited.Conclusion: In oral squamous cell carcinoma,silencing PDIA6 may inhibit the proliferation and migration of cancer cells through the MAPK/JNK,and it provides a theoretical basis for targeted therapy.