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
. 2018 Dec;53(6):2397-2408.
doi: 10.3892/ijo.2018.4579. Epub 2018 Oct 1.

Downregulation of CD147 induces malignant melanoma cell apoptosis via the regulation of IGFBP2 expression

Shuang Zhao  1 , Lisha Wu  2 , Yehong Kuang  1 , Juan Su  1 , Zhongling Luo  1 , Yan Wang  3 , Jinmao Li  1 , Jianglin Zhang  1 , Wangqing Chen  1 , Fangfang Li  1 , Yijing He  1 , Juan Tao  4 , Jianda Zhou  5 , Xiaowei Xu  6 , Cong Peng  1 , Xiang Chen  1
Affiliations

Downregulation of CD147 induces malignant melanoma cell apoptosis via the regulation of IGFBP2 expression

Shuang Zhao et al. Int J Oncol. 2018 Dec.

Erratum in

Abstract

Cluster of differentiation (CD)147, as a transmembrane glycoprotein, is highly expressed in a variety of tumors. Accumulating evidence has demonstrated that CD147 serves critical roles in tumor cell death and survival; however, the underlying mechanism requires further investigation. In the present study, it was revealed that CD147 knockdown significantly increased melanoma cell apoptosis. In addition, downregulation of CD147 reversed the malignant phenotype of melanoma, as demonstrated by the induction of tumor cell apoptosis in a xenograft mouse model. In addition, a human apoptosis antibody array was performed and 9 differentially expressed apoptosis-related proteins associated with CD147 were identified, including insulin-like growth factor-binding protein 2 (IGFBP2). Additionally, CD147 knockdown was observed to significantly decreased IGFBP2 expression at the mRNA and protein levels in melanoma cells. Providing that IGFBP2 is a downstream molecule in the phosphatase and tensin homolog (PTEN)/phosphoinositide 3-kinase (PI3K)/protein kinase B (AKT) signaling pathway, the effects of CD147 on this particular pathway were investigated. Interestingly, the expression of phosphorylated (p)-AKT and p‑mechanistic target of rapamycin was attenuated, whereas PTEN was markedly upregulated in CD147-underexpressing melanoma cells. Furthermore, application of a PI3K‑specific inhibitor also decreased IGFBP2 expression. Importantly, IGFBP2 was highly expressed in clinical tissues of melanoma compared with the control group, and its expression exhibited a positive association with CD147. The present study revealed that CD147 served a critical role in mediating the apoptosis of melanoma cells via IGFBP2 and the PTEN/PI3K/AKT signaling pathway. IGFBP2 and CD147 were observed to be overexpressed in clinical melanoma tissues; IGFBP2 was shown to be positively associated with CD147 expression, suggesting that CD147 may be considered as a potential therapeutic target for chemotherapy or prevention for in melanoma.

PubMed Disclaimer

Figures

Figure 1
Figure 1
Induction of downregulated CD147 expression in melanoma A375 cells. (A) Transfection efficiency of pLK0.1-sh-CD147 (C1 or C2) and the control sh-Mock vectors in melanoma A375 cells was determined via western blotting to confirm CD147 protein expression. Transfection with the sh-CD147-C2 vector notably inhibited CD147 protein expression in A375 cells. (B) Stable downregulation of CD147 in A375 sh-CD147-C1 and -C2, and the control sh-Mock-transfected cells was assessed via flow cytometry analysis. Transfection with pLK0.1-sh-Mock vector into A375 is referred to as Sh-Mock. C1 or C2, transfection with pLK0.1-sh-CD147 vector; CD147, cluster of differentiation 147; sh, short hairpin RNA.
Figure 2
Figure 2
Inhibition of CD147 promotes the apoptosis of melanoma cell lines. (A) Representative dot plots of Annexin V-FITC and PI staining. After A375 cells were stably transfected with sh-Mock (left) or sh-CD147 vector (right), apoptosis analysis was performed by flow cytometry. (B) Apoptotic rate of sh-CD147- and sh-Mock-transfected A375 cells. The percentage of apoptotic cells was significantly higher with decreased CD147 expression. (C and D) Protein expression levels of apoptosis-associated cle-PARP in sh-CD147- and sh-Mock-transfected A375 and SK-28 cells. (E) Transmission electron microscopy images of apoptosis of sh-CD147-transfected and sh-Mock-transfected A375 cells. Apoptotic bodies are indicated by arrows. n=3 for each experiment. Scale bar=2 μm. **P<0.01. CD147, cluster of differentiation 147; cle-PARP, cleaved poly (ADP-ribose) polymerase; FITC, fluorescein isothiocyanate; PI, propidium iodide; sh, short hairpin RNA.
Figure 3
Figure 3
IGFBP2 is an apoptosis-associated protein in sh-CD147 melanoma cells. (A) Cluster analysis of the expression of 43 apoptosis-associated proteins in sh-CD147- and sh-Mock-transfected A375 cells. Green presented downregulation and red indicated higher protein expression relative to the mean expression of all samples. (B) Relative mRNA expression of IGFBP2 in sh-Mock- and sh-CD147-transfected A375 cells. (C and D) Quantitation of the protein expression levels of IGFBP2 in sh-Mock- and sh-CD147-transfected A375 and SK-28 cells was performed via western blotting using ImageJ software. n=3 for each experiment. **P<0.01 vs. sh-Mock. Bcl-2, B-cell lymphoma 2; Bcl-w, Bcl-2-like protein 2; Bad, Bcl-2-associated agonist of cell death; BID, BH3 interacting-domain death agonist; BIM, Bcl-2-like protein 11; CD147, cluster of differentiation 147; CD40L, CD40 ligand; cIAP-2, cellular inhibitor of apoptosis 2; cytoC, cytochrome c; FasL, Fas ligand; HSP, heat shock protein; HTRA, HtrA serine peptidase; IGFBP2, insulin-like growth factor-binding protein 2; sh, short hairpin RNA; SMAC, second mitochondria-derived activator of caspases; TNF, tumor necrosis factor; sTNF-R2, soluble TNF receptor 2; TRAILR, TNF-related apoptosis-inducing ligand receptor; XIAP, X-linked inhibitor of apoptosis protein.
Figure 4
Figure 4
CD147 regulates IGFBP2 expression via the PTEN/PI3K/AKT signaling pathway. (A) Cell lysates of sh-CD147 and sh-Mock A375 and SK-MEL-28 cells were subjected to western blotting to detect p-AKT, p-mTOR and PTEN expression. Quantitation of (B) p-AKT, (C) p-mTOR and (D) PTEN expression was performed using ImageJ software. (E) Sh-Mock A375 and SK-MEL-28 cells were cultured with 50 μg/ml LY294002, a PI3K signaling pathway inhibitor, for 24 h. IGFBP2 and CD147 expression was detected by western blotting and (F) quantitative analysis of IGFBP2 expression was performed using ImageJ software. n=3 for each experiment. *P<0.05, **P<0.01 vs. sh-Mock or 0 μg/ml LY294002. AKT, protein kinase B; CD147, cluster of differentiation 147; m-TOR; p, phosphorylated; PI3K, phosphoinositide 3-kinase; PTEN, phosphatase and tensin homolog; sh, short hairpin RNA.
Figure 5
Figure 5
CD147 with IGFBP2 mediates the apoptosis of melanoma cells in vivo. (A) Tumor volume in the xenograft mouse model generated by inoculating melanoma sh-CD147 A375 and sh-Mock A375 cells. **P<0.05 or ***P<0.01 vs. sh-Mock. (B) Pathological characteristics of xenograft mouse tumors were observed by HE staining. Magnification, ×100. (C) Representative immunohistochemical staining for CD147, IGFBP2 or cle-PARP in serial sections of sh-CD147-injected xenograft mouse tumors or sh-Mock tumors. Magnification, ×100. The area positive for CD147, IGFBP2 and cleaved PARP- in sh-CD147 or sh-Mock cell-injected xenograft mouse tumors was determined by using Image-Pro Plus 6.0 software. (D) Representative TUNEL assay in sh-CD147-injected xenograft mouse tumors and sh-Mock tumors; the apoptosis-positive regions are indicated in brown-yellow. The histogram indicated the TUNEL-positive area (%) (right). Magnifications, ×100 or ×400. CD147, cluster of differentiation 147; cle-PARP, cleaved poly (ADP-ribose) polymerase; IGFBP2, insulin-like growth factor-binding protein 2; sh, short hairpin RNA; TUNEL, terminal deoxynucleotidyl-transferase-mediated dUTP nick end labeling.
Figure 6
Figure 6
CD147 and IGFBP2 overexpression in melanoma tissues. (A) Representative immunohistochemical staining of CD147- or IGFBP2-positive cells in nevus (n=15), PMM (n=128) and MMM (n=64) tissues. Magnification, ×100. (B) Comparison of the CD147- and IGFBP2-positive area (%) in nevus and melanoma microassay tissue. Semi-quantitation was performed using Image-Pro Plus 6.0 software. **P<0.05 MMM vs. PMM or nevus. MMM, metastatic malignant melanoma; PMM, primary malignant melanoma.
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

See all "Cited by" articles

References

    1. Zimmer L, Haydu LE, Menzies AM, Scolyer RA, Kefford RF, Thompson JF, Schadendorf D, Long GV. Incidence of new primary melanomas after diagnosis of stage III and IV melanoma. J Clin Oncol. 2014;32:816–823. doi: 10.1200/JCO.2013.49.5572. - DOI - PubMed
    1. Arnold M, Holterhues C, Hollestein LM, Coebergh JW, Nijsten T, Pukkala E, Holleczek B, Tryggvadóttir L, Comber H, Bento MJ, et al. Trends in incidence and predictions of cutaneous melanoma across Europe up to 2015. J Eur Acad Dermatol Venereol. 2014;28:1170–1178. doi: 10.1111/jdv.12236. - DOI - PubMed
    1. Yin H, Shao Y, Chen X. The effects of CD147 on the cell proliferation, apoptosis, invasion, and angiogenesis in glioma. Neurol Sci. 2017;38:129–136. doi: 10.1007/s10072-016-2727-2. - DOI - PubMed
    1. Kuang YH, Chen X, Su J, Wu LS, Liao LQ, Li D, Chen ZS, Kanekura T. RNA interference targeting the CD147 induces apoptosis of multi-drug resistant cancer cells related to XIAP depletion. Cancer Lett. 2009;276:189–195. doi: 10.1016/j.canlet.2008.11.010. - DOI - PubMed
    1. Xie W, Xie H, Liu F, Li W, Dan J, Mei Y, Dan L, Xiao X, Li J, Chen X. Propranolol induces apoptosis of human umbilical vein endothelial cells through downregulation of CD147. Br J Dermatol. 2013;168:739–748. doi: 10.1111/bjd.12127. - DOI - PubMed

MeSH terms