Additional file 1 of Histone lysine methyltransferase SMYD3 promotes oral squamous cell carcinoma tumorigenesis via H3K4me3-mediated HMGA2 transcription
Year: 2023
Type: dataset
Abstract: Additional file 1: Fig. S1. Identification of SMYD3 for diagnosis of OSCC. A–E ROC curve analyses of SMYD3 in TCGA, meta-GEO, TCGA, GSE37991, and GSE30784 datasets. AUC values are shown. Fig. S2. The DNA methylation and genomic mutation profile in the TCGA-OSCC dataset. A The correlation of SMYD3 expression and DNA methylation level in TCGA-OSCC cohort. B Value differences of DNA methylation probes in normaland tumortissues from TCGA-OSCC cohort. C The lollipop plot illustrates the differential distribution of somatic mutation in the TCGA-OSCC dataset for SMYD3. D, E ROC curve analyses of SMYD3 in qRT-PCR and IHC staining of collected samples, respectively. Ns, not significant, *P < 0.05, **P ≤ 0.01, and ***P ≤ 0.001. Fig. S3. High expression of SMYD3 indicates increased H3K4me3 modification and HMGA2 expression. A–F IHC images of high and low protein expression of SMYD3, H3K4me3 and HMGA2. Scale bars: 100 μm. Fig. S4. Biological function and pathway enrichment analysis. A The results of GO analysis of RNA-seq on two groups of CAL-27 transfected with NC and SMYD3 siRNA. B The results of KEGG analysis of RNA-seq on two groups of CAL-27 transfected with NC and SMYD3 siRNA. Fig. S5. SMYD3 facilitates OSCC cell stemness maintenance and proliferation in vitro and tumorigenesis in vivo. A, B SMYD3 mRNA and protein levels in CAL-27 and UM-SCC-1 cell lines. C SMYD3 mRNA levels in OSCC cells transfected with NC and SMYD3 siRNAs. D–G Quantitative statistical results of SMYD3 knockdown in vitro experiments. H SMYD3 mRNA levels in OSCC cells transfected with vector and SMYD3 plasmid. I–K Quantitative statistical results of SMYD3 overexpression in vitro experiments. L The protein expressions of SMYD3 and H3K4me3 were detected after transfection of CAL-27 cell line with SMYD3 plasmids. M, N SMYD3 mRNA and protein levels in CAL-27 transfected with shNC and shSMYD3. *P < 0.05, **P ≤ 0.01, and ***P ≤ 0.001. Fig. S6. BCI-121 suppresses OSCC cells stemness maintenance and proliferation. A H3K4me3 levels in OSCC cells under different concentrations of BCI-121. B, C The effect of different concentrations of BCI-121 on cell viability determined by CCK8 assays in CAL-27 and UM-SCC-1 cell lines. D SMYD3 mRNA levels in OSCC cells following BCI-121 treatment. E–H Quantitative statistical results of BCI-121 treatment in vitro experiments. Ns, not significant, *P < 0.05, **P ≤ 0.01, and ***P ≤ 0.001. Fig. S7. BCI-121 impedes the chemical-induced primary OSCC formation. A, B Representative H&E images of tongue lesions. Scale bars, 200 μm. C The number of clusters was determined using SOM clustering. Fig. S8. HMGA2 is upregulated in OSCC, and high expression of HMGA2 predicts a poor prognosis. A HMGA2 was overexpressed in OSCC samples from the meta-GEO dataset. Numbers in parentheses indicate the sample size. B Patients with high HMGA2 expression have a worse prognosis than those with low HMGA2 expression in the meta-GEO dataset. Numbers in parentheses indicate the sample size. C Quantitative result of qRT-PCR of HMGA2 in 20 paired adjacent normal and OSCC tissues. D The protein levels of HMGA2 in 12 pairs of OSCC tissuesand adjacent normal tissuesmeasured by Western blotting. E Images of IHC staining for HMGA2 in normal tissues and different histologic grades of OSCC tissues. Scale bars: 50 μm. Fig. S9. HMGA2 is a downstream target gene of SMYD3. A The SMYD3 binding sitein human HMGA2 promoter and the corresponding base mutation. B, C HMGA2 mRNA and protein levels in CAL-27 transfected with NC and HMGA2 siRNA. D Upon the completion of the ten-day tumorsphere formation assay, the proteins were extracted from the spheroid cells on the 10th day. The expression of SOX2 in SMYD3-upregulated cells was nullified following HMGA2 knockdown. E–H The tumorsphere formation and proliferation capacities of SMYD3-upregulated cells were abrogated by HMGA2 knockdown. Scale bars: 50 μm. *P < 0.05, **P ≤ 0.01, and ***P ≤ 0.001. Fig. S10. SMYD3 affects OSCC cell stemness maintenance and proliferation via HMGA2. A, B HMGA2 mRNA and protein levels in CAL-27 transfected with vector and HMGA2 plasmid. C–G The SOX2 expression, tumorsphere formation and proliferation capacities of HMGA2-upregulated cells were abrogated by SMYD3 knockdown. Scale bars: 50 μm. *P < 0.05, **P ≤ 0.01, and ***P ≤ 0.001
Source: Figshare
Authors: Yang, Zongcheng, Liu, Fen, Li, Zongkai, Liu, Nianping, Yao, Xinfeng, Zhou, Yu, Zhang, Liyu, Jiang, Pan, Liu, Honghong, Kong, Lingming, Lang, Chuandong, Xu, Xin, Jia, Jihui, Nakajima, Takahito, Gu, Wenchao, Zheng, Lixin, Zhang, Zhihong
Institutions: University of Science and Technology of China, Nanchang University, Xinjiang Medical University, Capital Medical University, First Affiliated Hospital of Xinjiang Medical University, Northwest Women's and Children's Hospital, Central South University, Yangtze University, University of California, San Francisco, Xiamen University, Shaoxing University, Jilin University, Chinese Academy of Sciences, Chinese Academy of Medical Sciences & Peking Union Medical College, Huazhong Agricultural University, Peking Union Medical College Hospital, Southern University of Science and Technology, Guangzhou Sport University, University of Jinan, Wuhan University, Fujian Institute of Research on the Structure of Matter, Qingdao Institute of Bioenergy and Bioprocess Technology, Arizona State University, Hebei GEO University, Southwest University of Science and Technology, Hebei Medical University, Harvard University, Shandong University, Shanghai Jiao Tong University, First Affiliated Hospital of Guangzhou Medical University, Nanfang Hospital, Second Affiliated Hospital of Nanjing Medical University, Xinxiang University, Huazhong University of Science and Technology, Zhejiang University, Nanjing Medical University, Xi'an Jiaotong University, University of Tsukuba, Gunma University, Changsha University, Affiliated Hospital of Southwest Medical University, Burning Rock Biotech (China), Anhui Provincial Hospital, Second Hospital of Tianjin Medical University, Jiangsu Province Hospital
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: green
OpenAlex ID: W4394544189