Long Noncoding RNA LINC00857 Promotes Proliferation, Migration, and Invasion of Colorectal Cancer Cell through miR-1306/Vimentin Axis

Abstract

Colorectal cancer is a commonly diagnosed cancer and the leading cause of cancer-related death which still increasing in many countries. The lack of biomarkers for early detection and clinic treatment results in high morbidity and mortality. The novel role of long noncoding RNA LINC00857 on cell proliferation migration and invasion was explored in this article. The expression level of LINC00857 in colorectal cancer tissue samples and cells was determined notably higher than normal tissue samples and cells. Silence LINC00857 can significantly inhibit colorectal cancer cell viability and metastasis in vitro. Moreover, LINC00857 depletion caused cell accumulation in the G0/G1 phase. In addition, we recognized the novel LINC00857-miR-1306-vimentin axis and demonstrated it by dual-luciferase reporter assay. And this signaling axis could be considered as the target for colorectal cancer treatment. In conclusion, LINC00857 can promote colorectal cancer progress by sponging miR-1306 and upregulate vimentin to accelerate the epithelial-mesenchymal transition process.

Figure 1

LINC00857 was determined highly expressed in colorectal cancer tissues and cell lines compared with normal tissues and cell lines. (a) Quantitative real-time PCR was performed to detect the expression level of LINC00857 in colorectal cancer tissues (n = 18) and the adjacent normal tissues (n = 18). ^∗P < 0.05. (b) LINC00857 expression level in colorectal cancer cell lines (RKO and HCT-116) and normal colorectal epithelial cell line (FHC) was determined by qRT-PCR. ^∗∗∗P < 0.001, ^∗∗∗∗P < 0.0001. (c) SiLINC00857 was used in LINC00857 knockdown in cell, and control group was transfected with siNC. The efficiency of knockdown was detected by qRT-PCR. ^∗∗∗∗P < 0.0001.

Figure 2

Silence LINC00857 can inhibit colorectal cancer cell proliferation in vitro. (a, b) The location of LINC00857 was detected by using PARIS kit and quantitative real-time PCR in HCT-116 and RKO cells. U6 was considered as nuclear control, and GAPDH was considered as cytoplasmic control for standardized. (c) Cell count kit-8 was applied to measure the viability of colorectal cancer cells between LINC00857 knockdown and control group in HCT-116 cell. ^∗∗P < 0.01, ^∗∗∗P < 0.001. (d) Cell count kit-8 was applied to measure the viability of colorectal cancer cells between LINC00857 knockdown and control group in RKO cell. ^∗∗P < 0.01.

Figure 3

LINC00857 knockdown result in colorectal cancer cell cycle arrested in G0/G1 phase. (a) Flow cytometry analysis of cell cycle was performed to analyze the cell cycle when LINC00857 was inhibited compared with the control group in RKO cell. ^∗∗P < 0.01, ^∗∗∗P < 0.001. (b) Flow cytometry analysis of cell cycle was performed to analyze the cell cycle when LINC00857 was inhibited compared with control group in HCT-116 cell. ^∗∗∗P < 0.001, ^∗∗∗∗P < 0.0001.

Figure 4

Colorectal cancer cell metastasis was decreased with LINC00857 depletion. Invasion and migration assays were performed to detect the colorectal cancer cell metastasis when LINC00857 was depleted compared with the control group. ^∗∗∗∗P < 0.0001.

Figure 5

miR-1306 was demonstrated can directly bind with LINC00857 and vimentin in cell. (a) miR-1306 was demonstrated can directly bind with LINC00857 in cell by dual-luciferase reporter assay. ^∗∗P < 0.01. (b) miR-1306 was demonstrated can directly bind with vimentin in cell by dual-luciferase reporter assay. ^∗∗P < 0.01. (c) Vimentin was overexpressed in RKO and HCT-116 cells with LINC00857 knockdown to rescue the expression level of vimentin. ^∗P < 0.05, ^∗∗∗P < 0.001. (d) Vimentin was overexpressed in RKO and HCT-116 cells with miR-1306 overexpressed to rescue the expression level of vimentin. ^∗P < 0.05, ^∗∗P < 0.01. (e–h) Overexpress vimentin in RKO and HCT-116 cells could reverse the inhibitory effect on cell viability from LINC00857 knockdown or miR-1306 overexpression by performing CCK-8 assay. ^∗P < 0.05, ^∗∗P < 0.01.