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Silencing of Long Non-coding RNA TTN-AS1 Inhibits Hepatocellular Carcinoma Progression by the MicroRNA-134/ITGB1 Axis

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Abstract

Background

Hepatocellular carcinoma (HCC) causes considerable mortality worldwide. Long non-coding RNA (lncRNA) TTN-AS1 has been recently identified as an oncogene in several cancers, but its role in HCC and the molecules remain largely unknown.

Aims

The study aims to probe the function of lncRNA TTN-AS1 in HCC progression and the molecules involved.

Methods

Differentially expressed lncRNAs between HCC and the adjacent normal tissues were analyzed using a microarray. TTN-AS1 expression in HCC and normal tissues and cells was determined. Targeting relationships between TTN-AS1 and miR-134 and between miR-134 and ITGB1 were validated. Artificial up-regulation or down-regulation of TTN-AS1, miR-134 and ITGB1 was introduced in HCC cells to probe their effects on the biological behaviors of HCC cells. Xenograft tumors were induced in nude mice for in vivo experiments.

Results

TTN-AS1 and ITGB1 were highly expressed, while miR-134 was poorly expressed in HCC tissues. TTN-AS1 enforced ITGB1 expression through sequestering miR-134. Silencing of TTN-AS1 or over-expression of miR-134 inhibited proliferation, invasion, migration, and resistance to death of Huh7 cells. Following miR-134 silencing, further down-regulation of ITGB1 suppressed the malignant behaviors of HUH7 cells. The similar results were reproduced in vivo.

Conclusion

The current study provided evidence that TTN-AS1 might promote HCC progression through sponging miR-134 and the following ITGB1 up-regulation. TTN-AS1 may serve as a potential target for HCC treatment.

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Abbreviations

ANOVA:

Analysis of variance

CCK-8:

Cell Counting Kit-8

ceRNA:

Competing endogenous RNA

FISH:

Fluorescence in situ hybridization

FITC:

Fluorescein isothiocyanate

GAPDH:

Glyceraldehyde-3-phosphate dehydrogenase

HBV/HCV:

Hepatitis B virus/hepatitis C virus

HCC:

Hepatocellular carcinoma

IgG:

Immunoglobulin G

ITGB1:

Integrin Beta-1

LC:

Liver cancer

LncRNAs:

Long non-coding RNAs

Mean ± SD:

Mean ± standard derivation

miRNAs:

MicroRNAs

MUT:

Mutant type

ncRNAs:

Non-coding RNAs

PBS:

Phosphate buffer saline

PI:

Propidium iodide

RIP:

RNA immunoprecipitation

TBST:

Tris-buffered saline + tween

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Authors and Affiliations

  1. Department of Hepatobiliary Surgery, Zaozhuang Municipal Hospital, No 41, Longtou Road, Shizhong District, Zaozhuang, 277101, Shandong, People’s Republic of China

    Yong Huang, Peng Chu & Guangjian Bao

Authors
  1. Yong Huang
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  2. Peng Chu
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  3. Guangjian Bao
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Contributions

YH contributed to the study design and manuscript preparation; YH and PC contributed to the definition of intellectual content and manuscript editing; PC is the guarantor of integrity of the entire study and contributed to the study concepts and manuscript review; GJB took charge of the acquisition and analysis of data and manuscript editing. All authors read and approved the final manuscript.

Corresponding author

Correspondence to Guangjian Bao.

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Conflict of interest

The authors declare no potential conflicts of interest.

Ethics statement

The study was ratified by the Clinical Ethical Committee of Zaozhuang Municipal Hospital. Signed informed consent was received from each eligible participant. All experimental procedures were conducted in line with the ethical guidelines for the study of experimental pain in conscious animals. Great attempts were made to minimize the usage and pain of animals.

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Huang, Y., Chu, P. & Bao, G. Silencing of Long Non-coding RNA TTN-AS1 Inhibits Hepatocellular Carcinoma Progression by the MicroRNA-134/ITGB1 Axis. Dig Dis Sci 66, 3916–3928 (2021). https://doi.org/10.1007/s10620-020-06737-x

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