Objective: To investigate the expression of long-chain non-coding RNAs DAPK1 and miR-182 in pancreatic cancer tissues and the role of DAPK1 and miR-182 in pancreatic cancer cell invasion and migration and its mechanism.
Methods: The expression of DAPK1 and miR-182 in different pancreatic cancer and adjacent tissues and different pancreatic cancer cells were detected by qPCR. Transwell invasion assay was used to detect the invasion ability of pancreatic cancer cells after DAPK1 expression. The changes of the migration ability of pancreatic cancer cells after DAPK1 expression were detected by scratch test. Double luciferase reporter gene was used to detect the interaction between DAPK1 and miR-182. Transwell invasion assay showed that miR-182 overexpression of DAPK1 could restore the invasive ability of pancreatic cancer cells. Western blot was used to detect the expression of ROCK-1/RhoA pathway protein after overexpression of miR-182 in DAPK1 cells. Phalloidin was used to label the cytoskeleton. The effect of miR-182 overexpression of DAPK1 on tumor size and volume of pancreatic cancer was detected by subcutaneous tumor formation in nude mice.
Results: The expression of DAPK1 was significantly decreased in pancreatic cancer tissues compared with adjacent tissues, and the expression of DAPK1 decreased gradually and the expression of miR-182 was opposite with the progression of tumor. DAPK1 was associated with pathological stage of pancreatic cancer and lymph node metastasis, while miR-182 was positively correlated. The expression level of DAPK1 in pancreatic cancer cell HS766T was the lowest. Overexpression of DAPK1 could inhibit the invasion and migration of pancreatic cancer cells. DAPK1 could bind specifically to 3'UTR of miR-182. Overexpression of miR-182 could restore the invasion and migration of pancreatic cancer cells after overexpression of DAPK1. The expression of ROCK-1/RhoA pathway protein was down-regulated by miR-182 after expression of DAPK1, and the expression of ROCK-1/RhoA pathway protein was restored. The expression of F-actin in LV5-DAPK1 group was significantly decreased, the formation of cell membrane wrinkles was significantly reduced, and the formation of pseudopodia was significantly reduced compared with LV5-DAPK1 + miR-182-mimic group. The tumor volume and weight of tumor-bearing mice in LV5-DAPK1 + miR-182-mimic group were significantly increased compared with LV5-DAPK1 group.
Conclusion: DAPK1 plays an important role in the development and progression of pancreatic cancer. DAPK1 can regulate the invasion and migration of pancreatic cancer cells through the regulation of miR-82 through ROCK-1/RhoA signaling pathway.
Keywords: ROCK-1/RhoA; invasion; miR-182; migration; pancreatic cancer.
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