J Korean Neurosurg Soc.  2024 May;67(3):364-375. 10.3340/jkns.2023.0155.

High Expression of KIFC1 in Glioma Correlates with Poor Prognosis

• Xue P
• Zheng J
• Li R
• Yan L
• Wang Z
• Jia Q
• Zhang L
• Li X

• KMID: 2554874
• DOI: http://doi.org/10.3340/jkns.2023.0155

Abstract

Objective
: Kinesin family member C1 (KIFC1), a non-essential kinesin-like motor protein, has been found to serve a crucial role in supernumerary centrosome clustering and the progression of several human cancer types. However, the role of KIFC1 in glioma has been rarely reported. Thus, the present study aimed to investigate the role of KIFC1 in glioma progression.
Methods
: Online bioinformatics analysis was performed to determine the association between KIFC1 expression and clinical outcomes in glioma. Immunohistochemical staining was conducted to analyze the expression levels of KIFC1 in glioma and normal brain tissues. Furthermore, KIFC1 expression was knocked in the glioma cell lines, U251 and U87MG, and the functional roles of KIFC1 in cell proliferation, invasion and migration were analyzed using cell multiplication, wound healing and Transwell invasion assays, respectively. The autophagic flux and expression levels matrix metalloproteinase-2 (MMP2) were also determined using imaging flow cytometry, western blotting and a gelation zymography assay.
Results
: The results revealed that KIFC1 expression levels were significantly upregulated in glioma tissues compared with normal brain tissues, and the expression levels were positively associated with tumor grade. Patients with glioma with low KIFC1 expression levels had a more favorable prognosis compared with patients with high KIFC1 expression levels. In vitro, KIFC1 knockdown not only inhibited the proliferation, migration and invasion of glioma cells, but also increased the autophagic flux and downregulated the expression levels of MMP2.
Conclusion
: Upregulation of KIFC1 expression may promote glioma progression and KIFC1 may serve as a potential prognostic biomarker and possible therapeutic target for glioma.

Keyword

Glioma; KIFC1; Autophagy; Matrix metalloproteinase-2

Figure

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  Fig. 1. Expression of KIFC1 via IHC staining in glioma and normal tissues. A : Representative IHC images of KIFC1 expression in normal and glioma tissues of different grades at 200× magnification. B : A semi-quantitative evaluation of KIFC1 expression levels was conducted based on the IHc findings outlined in the preceding section (A). The positive number of KIFC1 was normalized in the grade IV group, which was set at 1.0. bioinformatics datasets exhibited expression levels of KIFC1 in (C) gene enrichment pathway interactive analysis determined that KIFC1 mRNA expression levels were elevated in GBM and LGG tissues relative to normal tissues. D and E : by applying GlioVis online analysis, it was discovered that the expression levels of KIFC1 increased as the glioma grade rose. D : TCGA RNA sequencing database in addition to (E) CGGA RNA sequencing database. Applying bioinformatics, the prognostic impact of KIFC1 in glioma was determined. Information from the GlioVis data portal revealed that patients with glioma with elevated KIFC1 expression levels displayed shorter life expectancy (F and G). **p≤0.01. KIFC1 : kinesin family member C1, GBM : glioblastoma, LGG : low-grade gliomas, CGGA : the chinese glioma genome Atlas, TCGA : The cancer genome Atlas, IHC : immunohistochemical.

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  Fig. 2. Transfection efficiency of kinesin family member C1 (KIFC1) knockdown. Western blotting analysis demonstrated that the levels of KIFC1 expression in both (A and B) U251 and (C and D) U87Mg cell lines were substantially decreased after KIFC1 genetic knockdown. The expression levels of KIFC1 protein were normalized to the control (NC) group, with a value of 1.0. *p≤0.05, **p≤0.01.

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  Fig. 3. Autophagic flux enhanced by KIFC1 knockdown. A and B : Western blotting analysis revealed that the expression of LC3B increased following KIFC1 knockdown. C and D : Representative immunofluorescence imaging of KIFC1 and LC3B expression obtained from imaging flow cytometry demonstrated that LC3B expression was upregulated following KIFC1 knockdown. KIFC1 knockdown inhibited the glioma cell proliferation. MTT assay was conducted in U251 and U87Mg cells after KIFC1 knockdown for 7 days and the findings showed that the knockdown of KIFC1 inhibited glioma cell proliferation (E). *p≤0.05, **p≤0.01. NC : normalized to the control, KIFC1 : kinesin family member C1, LC3B : microtubule-associated protein 1 light chain 3 beta, MTT : cell multiplication, siRNA : small interfering RNA.

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  Fig. 4. Inhibition of the migration via KIFC1 knockdown. A and B : based on the wound healing assay, the number of invasive U251 and U87MG cells was significantly reduced in the KIFC1 knockdown group. Transwell assays indicated that the number of migrating U251 and U87MG cells in the KIFC1 knockdown group decreased significantly. C and D : Suppression of KIFC1 inhibits glioma cell invasion and suppresses MMP2 expression. E and F : gelatin zymography assay exhibited that MMP2 expression was decreased by KIFC1 knockdown. *p≤0.05, **p≤0.01. NC : normalized to the control, KIFC1 : kinesin family member C1, MMP2 : matrix metalloproteinase-2.

Reference

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