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. 2018 Dec 27;27(1):55-64.
doi: 10.3727/096504018X15201143705855. Epub 2018 Mar 9.

Knockdown of Urothelial Carcinoma-Associated 1 Suppressed Cell Growth and Migration Through Regulating miR-301a and CXCR4 in Osteosarcoma MHCC97 Cells

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Knockdown of Urothelial Carcinoma-Associated 1 Suppressed Cell Growth and Migration Through Regulating miR-301a and CXCR4 in Osteosarcoma MHCC97 Cells

Genglong Zhu et al. Oncol Res. .

Retraction in

Abstract

Liver cancer is one of the most common malignancies in the world and a leading cause of cancer-related mortality. Accumulating evidence has highlighted the critical role of long noncoding RNAs (lncRNAs) in various cancers. The present study aimed to explore the role of lncRNA urothelial carcinoma-associated 1 (UCA1) in cell growth and migration in MHCC97 cells and its underlying mechanism. First, we assessed the expression of UCA1 in MHCC97 and three other cell lines by RT-qPCR. Then the expression of UCA1, miR-301a, and CXCR4 in MHCC97 cells was altered by transient transfection. The effects of UCA1 and miR-301 on cell viability, migration, invasion, and apoptosis were assessed. The results revealed that UCA1 expression was relatively higher in MHCC97 cells than in MG63, hFOB1.19, and OS-732 cells. Knockdown of UCA1 reduced cell viability, inhibited migration and invasion, and promoted cell apoptosis. However, the effect of UCA1 knockdown on cell growth and migration was blocked by miR-301a overexpression, whose expression was regulated by UCA1. We also found that miR-301a positively regulated CXCR4 expression. CXCR4 inhibition reversed the effect of miR-301a overexpression on cell growth and migration. Moreover, miR-301a activated the Wnt/β-catenin and NF-κB pathways via regulating CXCR4. The present study demonstrated that UCA1 inhibition exerted an antigrowth and antimigration role in MHCC97 cells through regulating miR-301a and CXCR4 expression.

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Conflict of interest statement

The authors declare no conflicts of interest.

Figures

Figure 1
Figure 1
Urothelial carcinoma-associated 1 (UCA1) was highly expressed in MHCC97 cells. The expression of UCA1 in MHCC97, MG63, hFOB1.19, and OS-732 cells was measured by quantitative reverse transcription PCR (RT-qPCR) analysis.
Figure 2
Figure 2
Knockdown of UCA1 inhibited cell growth and migration in MHCC97 cells. MHCC97 cells were transfected with shNC, sh-UCA1#1, and sh-UCA1#2. (A) The efficiency of transfection was quantified by RT-qPCR analysis. (B) Cell viability was measured by trypan blue exclusion assay. (C) Cell migration and (D) invasion were detected by Transwell migration assay. (E) Apoptotic cells and (F) the levels of apoptosis-related proteins were measured by flow cytometry and Western blot, respectively. *p < 0.05, **p < 0.01, ***p < 0.001.
Figure 3
Figure 3
The expression of miR-301a was downregulated by UCA1 inhibition. MHCC97 cells were transfected with shNC and sh-UCA1#2. The expression of miR-301a was detected by RT-qPCR analysis. ***p < 0.001.
Figure 4
Figure 4
miR-301a overexpression blocked the effect of UCA1 on cell growth and migration in MHCC97 cells. MHCC97 cells were transfected with sh-UCA1 or shNC, or cotransfected with sh-UCA1 and miR-301a mimic or mimic control. (A) Cell viability, (B) cell migration, and (C) invasion were assessed by trypan blue exclusion assay and Transwell migration assay, respectively. (D) Apoptotic cells were quantified by flow cytometry assay. (E) Western blot was conducted to measure the expression of apoptosis-related core factors. *p < 0.05, **p < 0.01.
Figure 5
Figure 5
The expression of C-X-C chemokine receptor type 4 (CXCR4) was positively regulated by miR-301a. miR-301a mimic, miR-301a inhibitor, or their corresponding controls (mimic control and inhibitor control) were transfected into MHCC97 cells. (A) The mRNA and (B) protein expression of CXCR4 was determined by RT-qPCR analysis and Western blot analysis, respectively. *p < 0.05.
Figure 6
Figure 6
CXCR4 was involved in the regulatory effect of miR-301a on cell growth and migration. MHCC97 cells were transfected with sh-CXCR4 or sh-NC, or cotransfected with sh-CXCR4 and miR-301a mimic, or mimic control. The (A) mRNA and (B) protein expression levels of CXCR4 were tested by RT-qPCR analysis and Western blot analysis, respectively. (C) Cell viability, (D) cell migration, (E) invasion, and (F) apoptotic cells were assessed by trypan blue exclusion assay, Transwell migration assay, and flow cytometry assay, respectively. (G) The protein levels of apoptosis-related factors were measured by Western blot. *p < 0.05, **p < 0.01, ***p < 0.001.
Figure 7
Figure 7
miR-301a enhanced the activation of the Wnt/β-catenin and NF-κB signaling pathways through upregulating CXCR4. MHCC97 cells were transfected with miR-301a inhibitor, miR-301a mimic, or their corresponding controls (mimic control and inhibitor control), or cotransfected with miR-301a inhibitor and pEX-CXCR4. The protein expression of (A) Wnt/β-catenin- and (B) NF-κB signaling pathway-related core factors was measured by Western blot.

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References

    1. Wray CJ, Harvin JA, Silberfein EJ, Ko TC, Kao LS. Pilot prognostic model of extremely poor survival among high-risk hepatocellular carcinoma patients. Cancer 2012;118(24):6118–25. - PubMed
    1. Yeh YT, Chang CW, Wei RJ, Wang SN. Progesterone and related compounds in hepatocellular carcinoma: Basic and clinical aspects. BioMed Res Int. 2013;2013(12):290575. - PMC - PubMed
    1. Dhanasekaran R, Limaye A, Cabrera R. Hepatocellular carcinoma: Current trends in worldwide epidemiology, risk factors, diagnosis, and therapeutics. Hepat Med. 2012;4:19–37. - PMC - PubMed
    1. Yarmishyn AA, Kurochkin IV. Long noncoding RNAs: A potential novel class of cancer biomarkers. Front Genet. 2015;6:145. - PMC - PubMed
    1. Zhou M, Wang X, Shi H, Cheng L, Wang Z, Zhao H, Yang L, Sun J. Characterization of long non-coding RNA-associated ceRNA network to reveal potential prognostic lncRNA biomarkers in human ovarian cancer. Oncotarget 2016;7(11):12598–611. - PMC - PubMed

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