Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2022 Jan 15;14(1):68-85.
eCollection 2022.

The molecular mechanism of kinesin family member 2A (KIF2A) underlying non-small cell lung cancer: the effect of its knockdown on malignant behaviors, stemness, chemosensitivity, and potential regulated signaling pathways

Jiayan Chen  1   2 , Junmiao Wen  1   2 , Di Liu  1   2 , Xinyan Xu  1   2 , Min Fan  1   2 , Zhen Zhang  1   2
Affiliations

The molecular mechanism of kinesin family member 2A (KIF2A) underlying non-small cell lung cancer: the effect of its knockdown on malignant behaviors, stemness, chemosensitivity, and potential regulated signaling pathways

Jiayan Chen et al. Am J Transl Res. .

Abstract

Kinesin family member 2A (KIF2A) represents an oncogene in several cancers, however, its involvement in non-small cell lung cancer (NSCLC) is limitedly investigated. Therefore, the present study aimed to explore potential molecular mechanism of KIF2A knockdown in repressing NSCLC malignant behaviors. The effect of KIF2A knockdown on cell proliferation, apoptosis, invasion, epithelial-mesenchymal transition (EMT) markers, stemness, chemosensitivity was detected after transfecting KIF2A short hairpin RNA (ShRNA) plasmids into A549 and NCI-H1975 cells. Moreover, KIF2A knockdown mediated signaling pathways were analyzed by RNA sequencing (RNA-seq), and then validated by western blot assay. Both KIF2A mRNA and protein expressions were increased in A549, NCI-H650, NCI-H358, NCI-H2106, NCI-H1299, NCI-H1650 and NCI-H1975 cells compared with BEAS-2B cells. KIF2A knockdown inhibited proliferation, invasion, EMT, stemness, but enhanced chemosensitivity to cisplatin and paclitaxel in both A549 and NCI-H1975 cells. Meanwhile, it only promoted apoptosis in NCI-H1975 cells but not in A549 cells. Moreover, after KIF2A knocking down, RNA-seq data indicated that 356 accordant differentially expressed genes (DEGs) in both A549 and NCI-H1975 cells, and these DEGs were enriched in PI3K-Akt, Wnt and Notch signaling pathways. Further western blot disclosed that KIF2A knockdown indeed inactivated PI3K-Akt, Wnt and Notch signaling pathways in both A549 and NCI-H1975 cells. In conclusion, KIF2A knockdown suppresses NSCLC cell malignant behaviors, EMT and stemness, but enhances chemosensitivity via inactivating PI3K-Akt, Wnt, and Notch signaling pathways, which proposes it as a potential therapeutic target for NSCLC treatment.

Keywords: Kinesin family member 2A; RNA-sequencing; chemosensitivity; malignant behaviors; non-small cell lung cancer.

PubMed Disclaimer

Conflict of interest statement

None.

Figures

Figure 1
Figure 1
Comparison of KIF2A expression. Comparison of KIF2A mRNA (A) and protein (B) expressions between human NSCLC cell lines (A549, NCI-H650, NCI-H358, NCI-H2106, NCI-H1299, NCI-H1650, NCI-H1975 cell lines) and normal lung epithelial cell line (BEAS-2B cell line). KIF2A, kinesin family member 2A; NSCLC, non-small cell lung cancer.
Figure 2
Figure 2
KIF2A expression of transfected NSCLC cells. After transfecting three KIF2A ShRNA plasmids into A549 (A, B) and NCI-H1975 (C, D) cells, comparisons of KIF2A mRNA and protein expressions between Sh-KIF2A1, Sh-KIF2A2, Sh-KIF2A3 groups and Sh-NC group, respectively. KIF2A, kinesin family member 2A; NSCLC, non-small cell lung cancer; Sh, short hairpin RNA; NC, negative control.
Figure 3
Figure 3
Comparison of cell proliferation and apoptosis. Comparison of cell proliferation (A), cell apoptosis (B, C) between Sh-KIF2A group and Sh-NC group in A549 cells. Comparison of cell proliferation (D), cell apoptosis (E, F) between Sh-KIF2A group and Sh-NC group in NCI-H1975 cells. KIF2A, kinesin family member 2A; Sh, short hairpin RNA; NC, negative control; OD, optical density; CCK-8, cell counting kit-8.
Figure 4
Figure 4
Comparison of cell invasion and EMT markers. Comparison of invasive cells count (A, B), expressions of EMT markers (C) between Sh-KIF2A group and Sh-NC group in A549 cells. Comparison of invasive cells count (D, E), expressions of EMT markers (F) between Sh-KIF2A group and Sh-NC group in NCI-H1975 cells. KIF2A, kinesin family member 2A; Sh, short hairpin RNA; NC, negative control.
Figure 5
Figure 5
Comparison of stemness. Comparison of sphere formation ability (A), and CD133+ cells proportion (B, C) between Sh-KIF2A group and Sh-NC group in A549 cells. Comparison of sphere formation ability (D), and CD133+ cells proportion (E, F) between Sh-KIF2A group and Sh-NC group in NCI-H1975 cells. KIF2A, kinesin family member 2A; Sh, short hairpin RNA; NC, negative control.
Figure 6
Figure 6
Comparison of chemosensitivity. Comparison of relative cell viability between Sh-KIF2A group and Sh-NC group treated with different concentration of cisplatin (A) and paclitaxel (B) in A549 cells. Comparison of relative cell viability between Sh-KIF2A group and Sh-NC group treated with different concentration of cisplatin (C) and paclitaxel (D) in NCI-H1975 cells. KIF2A, kinesin family member 2A; Sh, short hairpin RNA; NC, negative control.
Figure 7
Figure 7
PCA plots and heatmap analysis. PCA plot (A) and heatmap analysis (B) for mRNA profiles by KIF2A knockdown in A549 cells. PCA plot (C) and heatmap analysis (D) for DEGs by KIF2A knockdown in NCI-H1975 cells. KIF2A, kinesin family member 2A; Sh, short hairpin RNA; NC, negative control; PCA, principal component analysis.
Figure 8
Figure 8
Volcano plot and KEGG enrichment analysis. Volcano plot (A), KEGG enrichment analysis (B) for DEGs by KIF2A knockdown in A549 cells. Volcano plot (C), KEGG enrichment analysis (D) for DEGs by KIF2A knockdown in NCI-H1975 cells. KIF2A, kinesin family member 2A; Sh, short hairpin RNA; NC, negative control; DEGs, differentially expression genes.
Figure 9
Figure 9
Venn diagram and KEGG enrichment analysis for accordant DEGs. Venn diagram analysis for the accordant DEGs by KIF2A knockdown in both A549 and NCI-H1975 cells (A). KEGG enrichment analysis for these accordant DEGs (B). KIF2A, kinesin family member 2A; Sh, short hairpin RNA; NC, negative control; DEGs, differentially expression genes.
Figure 10
Figure 10
Validation of 3 potential signaling pathway regulated by KIF2A knockdown. Validation of KIF2A knockdown on PI3K-Akt, Wnt and Notch signaling pathways in A549 (A) and NCI-H1975 (B) cells. KIF2A, kinesin family member 2A; Sh, short hairpin RNA; NC, negative control.
See this image and copyright information in PMC

References

    1. Osmani L, Askin F, Gabrielson E, Li QK. Current WHO guidelines and the critical role of immunohistochemical markers in the subclassification of non-small cell lung carcinoma (NSCLC): moving from targeted therapy to immunotherapy. Semin Cancer Biol. 2018;52:103–109. - PMC - PubMed
    1. Nagasaka M, Gadgeel SM. Role of chemotherapy and targeted therapy in early-stage non-small cell lung cancer. Expert Rev Anticancer Ther. 2018;18:63–70. - PMC - PubMed
    1. Arbour KC, Riely GJ. Systemic therapy for locally advanced and metastatic non-small cell lung cancer: a review. JAMA. 2019;322:764–774. - PubMed
    1. Valentino F, Borra G, Allione P, Rossi L. Emerging targets in advanced non-small-cell lung cancer. Future Oncol. 2018;14:61–72. - PubMed
    1. Ettinger DS, Aisner DL, Wood DE, Akerley W, Bauman J, Chang JY, Chirieac LR, D’Amico TA, Dilling TJ, Dobelbower M, Govindan R, Gubens MA, Hennon M, Horn L, Lackner RP, Lanuti M, Leal TA, Lilenbaum R, Lin J, Loo BW, Martins R, Otterson GA, Patel SP, Reckamp K, Riely GJ, Schild SE, Shapiro TA, Stevenson J, Swanson SJ, Tauer K, Yang SC, Gregory K, Hughes M. NCCN guidelines insights: non-small cell lung cancer, version 5.2018. J Natl Compr Canc Netw. 2018;16:807–821. - PubMed

LinkOut - more resources