Limb-Bud and Heart Attenuates Growth and Invasion of Human Lung Adenocarcinoma Cells and Predicts Survival Outcome

Abstract

Background/Aims: The transcription cofactor limb-bud and heart (LBH) is involved in embryonic development. However, its role in human lung cancer, especially lung adenocarcinoma (LUAD), remains unclear. Methods: A public database and tissue microarray (TMA) were used to compare differences in LBH expression and its relationship with clinical characteristics. Tissue from an additional 70 LUAD patients with follow-up records was used to explore the correlation of LBH expression with prognosis. Cellular and molecular studies validated the role of LBH in LUAD growth and invasion. Results: LBH was significantly down-regulated in lung cancer tissue samples and was correlated with the prognosis and clinical characteristics of lung cancer patients based on a public database and TMA. Survival analysis revealed that LBH-negative expression was associated with poor overall survival of LUAD patients (P = 0.021). Cox regression analysis showed that LBH expression status was a favorable independent prognostic factor (hazard ratio = 0.120, 95% confidence interval = 0.016–0.894, P = 0.039). LBH knockdown accelerated LUAD cell proliferation, migration, and invasion. Furthermore, bioinformatics analysis indicated that LBH was significantly related to the cell adhesion pathway. Western blot analysis confirmed that LBH could regulate the expression of integrin family members (integrin-α1, integrin-α2, integrin-α4, integrin-αv, and integrin-β4). Conclusion: Our data suggest that LBH plays an important role in lung cancer. Importantly, LBH is an independent prognostic factor in LUAD and can attenuate cell growth and invasion. LBH may be a potential prognostic biomarker in LUAD patients.

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“…In order to detect the long‐term effects of the expression of LRRN1 on cell viability, 500 cells/well were added to 12‐well culture plates for 2 weeks. The cells were fixed with methanol and then stained with 0.5% crystal violet, and the colonies were counted using Image‐Pro Plus …”

Knockdown of G‐protein‐signaling modulator 2 promotes metastasis of non‐small‐cell lung cancer by inducing the expression of Snail

“…Immunohistochemical staining was carried out as described previously. 17 Staining was carried out with an anti-human GPSM2 rabbit Ab that was used at a dilution of 1:100 (ab84571; Abcam); PBS was used as a negative control. Each section was evaluated and scored independently by 2 pathologists.…”

“…RNA was isolated and reverse transcribed as previously described. 17 The primer sequences for amplification were as follows: forward 5′-AGCCAGTCGGTACTTAGCCA-3′ and reverse 5′-TTGTGGTAGCAGGTGGTGGA-3′ for GPSM2, and forward 5′-GGTGAAGGTCGGAGTCAACGG-3′ and reverse 5′-GAGGTCAATGAAGGGGTCATTG-3′ for the 18S control.…”

“…Western blot analysis was carried out as previously described. 17 The following primary Abs were used: GPSM2 rabbit polyclonal Ab…”

Localization of GPSM2 in the Nucleus of Invasive Breast Cancer Cells Indicates a Poor Prognosis

“…IHC staining was performed as described previously (19). Antihuman GPSM2 rabbit antibody was used at a dilution of 1:100 (ab84571; Abcam, Cambridge, UK); Anti-human DYNC1I1 mouse antibody was used at a dilution of 1:100 (ab23905; Abcam, Cambridge, UK); phosphate-buffered saline was used as a negative control.…”