Abnormal CBLL1 methylation and mRNA expression in pan-cancerous tissues
In SMART database, we found 17 CBLL1 gene probes and abnormal CBLL1 methylation levels in pan-cancerous tissues (Fig. 1 and Fig. S1-13). Probe cg10861925 organizes abnormalities in BLCA, BRCA, HNSC, KIRC, LUSC, PAAD, PGAD, READ, SARC and THCA (Fig. 1A); probe cg02952735 organizes abnormalities in BLCA, BRCA, HNSC, KIRC, KIRP, LIHC, LUAD, LUSC, PAAD, PRAD, READ, SARC and THCA (Fig. 1B); probe cg03443922 organizes abnormalities in BLCA, BRCA, COAD, ESCA, HNSC, KIRC, LIHC, LUSC, PRAD, READ and READ (Fig. 1C). Probe cg19318403 organizes abnormalities in BLCA, BRCA, ESCA, KIRC, KIRP, LIHC, LUAD, LUSC, PCPG, PRAD and THCA (Fig. 1D); probe cg06724108 organizes abnormalities in KIRC, LUSC, SARC and SKCM (Fig. S1); probe cg07614064 organizes abnormalities in BLCA, COAD, KIRC, LUAD, LUSC, READ, THCA and UCEC (Fig. S2); probes cg01331191 organizes abnormalities in BLCA, COAD, HNSC, KIRC, KIRP, LIHC, LUAD, PRAD and UCEC (Fig. S3). Probe cg03147542 organizes abnormalities in BRCA, CHOL, COAD, ESCA, HNSC, PAAD and SARC (Fig. S4); probe cg26385438 organizes abnormalities in BRCA, HNSC, KIRC, LIHC, LUSC, PAAD, PRAD and SARC (Fig. S5); probe cg25183958 organizes abnormalities in BLCA, LIHC, LUSC, PAAD, PRAD, SARC, THCA and UCEC (Fig. S6); probe cg15056119 organizes abnormalities in LIHC, LUSC, PAAD, PRAD, READ, SARC and UCEC (Fig. S7). Probe cg04433990 is abnormal in BLCA, BRCA, HNSC, KIRC, LUSC, and SARC (Fig. S8); probe cg20263098 is abnormal in BRCA, HNSC, KIRC, LIHC, LUSC, PAAD, PRAD, READ and SARC (Fig. S9); probe cg21810311 is abnormal in BLCA, COAD, KIRC, KIRP, LUSC, SARC and SKCM (Fig. S10); probe cg00449210 organizes abnormalities in KIRP, LIHC, SARC and THCA (Fig. S11); probe cg27396403 organizes abnormalities in BRCA, COAD, KIRC, LIHC, LUSC, LUSC and PRAD (Fig. S12). Probe cg17158101 was abnormal in BLCA, KIRP, LIHC, LUSC, PAAD, PCPG, PRAD, READ, SARC and STAD (Fig. S13). In addition, in the UCSC database, we found abnormal expression of CBLL1 in a variety of tumors (Fig. S14 and Fig. 2). In detail, CBLL1 is highly expressed in CHOL, COAD, ESCA, GBM, KICH, LIHC, LUSC, HNSC, READ and STAD tissues (Fig. 2A-J), and poorly expressed in THCA and UCEC tissues (Fig. 2K-L).
The expression of CBLL1 is related to the age, race, clinical stage and therapeutic effect of patients with pan-cancer
After collating and analyzing the clinicopathological features of 33 cancer patients, it was found that the expression level of CBLL1 was correlated with the age of BRCA, CHOL, KIRP, LIHC and PAAD patients (Fig. S15), the expression level of CBLL1 mRNA was correlated with the race of patients with BLCA, BRCA, GBM, KIRC, LIHC and THYM (Fig. S16), and the expression level of CBLL1 mRNA was correlated with the clinical stage of patients with ACC, BRCA, KIRC, LIHC, PAAD, STAD, TGCT and THCA (Fig. 3). The expression level of CBLL1 mRNA was related to the therapeutic effect of patients with BLCA, KIRC, KIRP, LGG, PRAD and UCEC (Fig. S17).
The expression of CBLL1 is related to the prognosis of patients with pan-cancer
Survival analysis of clinical information of 33 kinds of cancer patients showed that the expression level of CBLL1 mRNA was correlated with OS in patients with KIRC (Fig. 4A), the expression level of CBLL1 mRNA was correlated with DSS in patients with KIRC, LUSC, THCA and THYM (Fig. 4B-E), the expression level of CBLL1 mRNA was correlated with DFI in patients with MESO, PRAD and STAD (Fig. 4F-H), and the expression level of CBLL1 mRNA was correlated with PFI in patients with KIRC, PRAD and UVM (Fig. 7I-K). Univariate COX regression analysis found that CBLL1 mRNA expression level was a risk factor for OS in patients with KICH, KIRC, LAML and THYM (Fig. S18A);DSS in patients with KIRC, PCPG and THYM (Fig. S18B); DFI in patients with OV, PRAD and STAD (Fig. S18C); PFI in patients with GBM, KIRC, OV, THYM and UVM (Fig. S18D).
The expression level of CBLL1 is associated with TMB and MSI in patients with pan-cancer
The mutation load of pan-cancer tumor was calculated based on the expression data of pan-cancer mRNA in UCSC database, and the correlation between the expression level of CBLL1 mRNA and TMB of pan-cancer patients was analyzed. In other words, the expression level of CBLL1 mRNA was correlated with TMB in patients with BLCA, BRCA, COAD, LAML, LGG, LUAD, LUSC, SARC, STAD, THCA, THYM and UVM (Fig. S19A and Table 1); The expression level of CBLL1 mRNA was correlated with MSI in patients with pan-cancer. In detail, the expression level of CBLL1 mRNA was associated with ACC, BRCA, CESC, COAD, DLBC, HNSC, PRAD, READ, SARC, STAD, TGCT, THCA and MSI in patients with UCEC (Fig. S19B and Table 2).
Table 1
The expression level of CCBL1 is related to TMB in patients with pan-cancer.
Cancer | cor | P | Cancer | cor | P | Cancer | cor | P |
ACC | 0.054142902 | 0.635564064 | KIRC | -0.102232312 | 0.062798785 | PRAD | -0.008189613 | 0.857674661 |
BLCA | 0.148662078 | 0.002609396 | KIRP | -0.029322985 | 0.626390391 | READ | 0.034712864 | 0.692734917 |
BRCA | -0.109007953 | 0.000654847 | LAML | 0.270586236 | 0.031961519 | SARC | 0.140875647 | 0.030862194 |
CESC | 0.095841162 | 0.105781572 | LGG | 0.234959513 | 1.03E-07 | SKCM | 0.038816735 | 0.403661151 |
CHOL | 0.021641127 | 0.900302869 | LIHC | -0.008575317 | 0.871372482 | STAD | 0.218241875 | 2.41E-05 |
COAD | -0.228484562 | 4.36E-06 | LUAD | 0.231940191 | 1.43E-07 | TGCT | -0.039723537 | 0.635227514 |
DLBC | -0.199146515 | 0.236406449 | LUSC | 0.103243922 | 0.022549494 | THCA | -0.147243369 | 0.001187156 |
ESCA | -0.070773267 | 0.373833529 | MESO | 0.071290726 | 0.532403545 | THYM | -0.203247682 | 0.027959796 |
GBM | 0.096803132 | 0.241829324 | OV | 0.069487861 | 0.253401475 | UCEC | 0.052933772 | 0.225963196 |
HNSC | 0.088138064 | 0.050721582 | PAAD | 0.064774996 | 0.429420079 | UCS | -0.116530763 | 0.392392087 |
KICH | 0.082648833 | 0.512771489 | PCPG | 0.05657479 | 0.454493292 | UVM | -0.236577168 | 0.03461792 |
Note: cor, correlation coefficient. |
Table 2
The expression level of CCBL1 is related to MSI in patients with pan-cancer.
Cancer | cor | P | Cancer | cor | P | Cancer | cor | P |
ACC | 0.331783493 | 0.002816794 | KIRC | -0.071313799 | 0.192903946 | PRAD | -0.14023156 | 0.001762789 |
BLCA | 0.052697193 | 0.288279558 | KIRP | -0.043347334 | 0.466054376 | READ | 0.196212708 | 0.015405561 |
BRCA | -0.080131522 | 0.010053698 | LAML | 0.102531803 | 0.405397976 | SARC | 0.151246453 | 0.01605443 |
CESC | 0.128669674 | 0.025347158 | LGG | -0.067684434 | 0.127628875 | SKCM | -0.006347044 | 0.891076845 |
CHOL | 0.107850708 | 0.529849386 | LIHC | -0.00963497 | 0.853654122 | STAD | 0.115846826 | 0.025063683 |
COAD | -0.205064265 | 1.95E-05 | LUAD | 0.057166114 | 0.196999196 | TGCT | -0.199563168 | 0.014350286 |
DLBC | -0.677595334 | 1.23E-07 | LUSC | -0.028070043 | 0.534075678 | THCA | -0.094575424 | 0.036170301 |
ESCA | 0.024476121 | 0.75867615 | MESO | 0.210823638 | 0.057274624 | THYM | -0.033674092 | 0.717349781 |
GBM | 0.079716687 | 0.330561054 | OV | 0.099127277 | 0.102818564 | UCEC | 0.102287651 | 0.017632145 |
HNSC | -0.106990995 | 0.017142546 | PAAD | -0.033739987 | 0.657573586 | UCS | 0.095697051 | 0.482933666 |
KICH | 0.105036553 | 0.405009768 | PCPG | 0.043598971 | 0.563358916 | UVM | 0.179750916 | 0.110608901 |
Note: cor, correlation coefficient. |
The expression level of CBLL1 mRNA is related to the microenvironment of pan-cancer
The expression level of CBLL1 mRNA was correlated with tumor microenvironment (Fig. 5, Fig. S20 and Table 3). Figure 5 shows the correlation between CBLL1 mRNA expression level and tumor stromal cells in the top 9 P values. In detail, the expression level of CBLL1 was correlated with tumor stromal cells such as TGCT, LGG, SARC, GBM, LUSC, BLCA, BRCA, THCA, PCPG and so on (Fig. 5 and Table 3). Figure S20 shows the correlation between CBLL1 mRNA expression level and tumor immune cells in the top 9 P values. In detail, the expression level of CBLL1 was correlated with tumor immune cells such as BRCA, LGG, THCA, GBM, LUSC, SARC, UCEC, PCPG, LIHC and so on (Fig. S20 and Table 3).
Table 3
The expression level of CBLL1 mRNA is related to the immune infiltration of pan-cancerous tumors.
Cancer type | StromalScore | ImmuneScore |
ACC | 0.124603384 | 0.010137825 |
BLCA | 3.84E-05 | 0.001271417 |
BRCA | 3.89E-05 | 9.56E-11 |
CESC | 0.393921421 | 0.002109213 |
CHOL | 0.614788213 | 0.40869677 |
COAD | 0.963871374 | 0.001587956 |
DLBC | 0.234894177 | 0.952807531 |
ESCA | 0.627477696 | 0.515859015 |
GBM | 3.19E-08 | 1.60E-07 |
HNSC | 0.749378092 | 0.262083978 |
KICH | 0.248132174 | 0.134449688 |
KIRC | 0.083590093 | 0.001019052 |
KIRP | 0.512345762 | 0.002132654 |
LAML | 0.664580379 | 0.027041677 |
LGG | 2.10E-11 | 5.00E-09 |
LIHC | 0.137828166 | 8.44E-06 |
LUAD | 0.187731048 | 0.500395871 |
LUSC | 3.08E-05 | 0.00021417 |
MESO | 0.03062979 | 0.055479795 |
OV | 0.024490708 | 0.004501272 |
PAAD | 0.516894301 | 0.99423637 |
PCPG | 0.000883697 | 5.41E-06 |
PRAD | 0.62106203 | 0.021780091 |
READ | 0.320999231 | 0.089231316 |
SARC | 3.89E-11 | 6.05E-07 |
SKCM | 0.048128866 | 0.001705958 |
STAD | 0.016515899 | 0.074448303 |
TGCT | 0 | 0.002831903 |
THCA | 0.000341586 | 3.17E-08 |
THYM | 0.000961122 | 0.16042945 |
UCEC | 0.202244891 | 8.63E-07 |
UCS | 0.716896263 | 0.208104708 |
UVM | 0.524138205 | 0.511113834 |
The expression level of CBLL1 is related to the immune cells of pan-cancerous tumors
Figure 6 and figure S21 show the correlation between the expression level of CBLL1 mRNA and the tumor immune infiltrating cells with the lowest P value. The expression level of CBLL1 is correlated with BLCA (T cells follicular helper), BRCA (Tcells CD4 memory resting), COAD (B cells memory), GBM (T cells gamma delta), HNSC (B cell snaive), KICH (T cells CD4 memory resting), KIRC (T cells regulatory,Tregs), LAML (Monocytes), LIHC (T cells CD8), LUAD (T cells CD4 memory activated), LUSC (Neutrophils), OV (T cells CD8.), PRAD (T cells CD4 memory resting), READ (Macrophages M1), SARC (Macrophages M2), SKCM (Tregs), STAD (Tregs), TGCT (B cells naive), THCA (Dendritic cells activated), THYM (Macrophages M0), UCEC (T cells CD4 memory resting), UVM (Monocytes) and KIPR (T cells CD8)( Fig. 6 and Fig. S21; Supplementary table S1). In addition, the expression level of CBLL1 was associated with various immune infiltrating cells in pan-cancer. For example, the expression level of CBLL1 was significantly correlated with immune cells such as BRCA B cells naive, B cells memory, T cells CD8, T cells CD4 memory activated, Tregs; HNSC B cells naive, B cells memory, T cells CD4 memory resting, NK cells activated; PRAD B cells naive, B cells memory, Plasma cells, T cells CD8, T cells CD4 memory resting (P < 0.05, Supplementary table S1).
The expression level of CBLL1 is related to immunomodulators, checkpoints and receptor molecules in pan-cancerous tumors
To further explore the relationship between CBLL1 mRNA expression and tumor immune markers, in order to understand the role of CBLL1 in pan-cancer immune escape. immunomodulators include immunostimulators, immunoinhibitors and MHC molecules. We found that the expression level of CBLL1 was associated with pan-cancer immunomodulators (Fig. 7A-C). For example, the expression level of CBLL1 was correlated with BLCA immunostimulators CD160, ADORA2A, TGFBR1, IL10RB and BTLA, with BRCA immunostimulators LGALS9, IL10RB, CD160, KDR, TGFBR1, etc., and with CESC immunostimulators KDR, ADORA2A, LAG3, LGALS9 and IL10RB (P < 0.05, Fig. 7A). The expression level of CBLL1 was correlated with BLCA immunoinhibitors TNFRSF4, TNFRSF14, CD276TNFRSF18, TNFSF9, TNFRSF25, BRCA immunoinhibitors TNFRSF4, TNFRSF14, ULBP1, MICB, TNFSF18, and CESC immunoinhibitors TNFSF9, TNFSF18, NT5ETNFRSF4, TNFSF13 (P < 0.05, Fig. 7B). The expression level of CBLL1 was significantly correlated with BLCA MHC molecules HLA-A, HLA-E, HLA-F, TAPBP and HLA-DPB1, BRCA MHC molecules HLA-A, HLA-F, HLA-EHLA-C, HLA-G, HLA-B, and CESC MHC molecules HLA-F, HLA-C, HLA-B, HLA-A, TAPBP (P < 0.05, Fig. 7C). In addition, the expression level of CBLL1 was associated with pancancerous immune checkpoint molecules (Fig. 7D). For example, the expression level of CBLL1 was correlated with BLCA checkpoint molecules CCL26, CCL14, CCL23, CCL21, CCL28; BRCA checkpoint molecules CCL26, CCL3, CCL17, CCL23, CX3CL1, CXCL2; CESC checkpoint molecules CXCL2, CXCL3, CCL3, CXCL16, CCL13 (P < 0.05, Fig. 7D). In addition, the expression level of CBLL1 was associated with receptor molecules (Fig. S22 and Table 4). For example, the expression level of CBLL1 is related to BLCA receptor molecules CCR9, CCR8, CCR4, CCR7 and CXCR5; BRCA receptor molecules CCR10, CCR8, CCR9, CCR4, CXCR3, etc; CESC molecules CCR8, XCR1, CCR2, CCR6, CCR4, etc (Table 4).
Table 4
The expression level of CBLL1 is related to tumor immune receptor molecules.
Receptor | BLCA | BRCA | CESC |
CCR1 | 0.937418488 | 0.298791645 | 0.054925253 |
CCR2 | 0.606569164 | 0.002536219 | 0.009550921 |
CCR3 | 0.12648539 | 0.648938001 | 0.123018037 |
CCR4 | 0.010496864 | 0.001147277 | 0.014677045 |
CCR5 | 0.819332104 | 0.243023525 | 0.406713675 |
CCR6 | 0.053292813 | 0.003573498 | 0.010188994 |
CCR7 | 0.033661703 | 0.421512886 | 0.894509139 |
CCR8 | 0.000136705 | 3.35E-07 | 4.25E-05 |
CCR9 | 4.94E-06 | 0.000473587 | 0.866908606 |
CCR10 | 0.102926437 | 2.24E-21 | 0.149478695 |
CXCR1 | 0.219670706 | 0.923072873 | 0.021085763 |
CXCR2 | 0.129164084 | 0.114858105 | 0.514219098 |
CXCR3 | 0.510329501 | 0.00186531 | 0.77737012 |
CXCR5 | 0.044729072 | 0.576671819 | 0.018422254 |
CXCR6 | 0.132770807 | 0.25368142 | 0.954407026 |
XCR1 | 0.677307888 | 0.21604114 | 0.001885185 |
CX3CR1 | 0.233819871 | 0.002982067 | 0.507290776 |
The possible mechanism of the involvement of CBLL1 in the progression of pan-cancer
The expression of CBLL1 mRNA is related to the prognosis and immunity of patients with pan-cancer. Therefore, we use GSEA to analyze the possible mechanism of CBLL1 involved in tumor progression. The results showed that CBLL1 was involved in the development of pan-cancer through a variety of signal pathways (Fig. 8, Fig. S23 and Supplementary table S2-S34). For example, CBLL1 may participate in ACC progress through B cell receptor singaling pathway, mRNA cis splicing via spliceosome, immunoglobulin receptor binding, olfactory receptor activity, membrane disruption in other organism, tRNA threonylcarbamoyladenosine metabolic process, etc. (Fig. 8A and Supplementary table S2). CBLL1 may participate in the progress of BLCA through gene singaling, mRNA binding, regulation of cellualr amide metabolic process, endothelial cell migration, regulation of epithelial cell migration, etc. (Fig. 8B and Supplementary table S3). CBLL1 may participate in the progress of BRCA through positive regulation of cell cycle phase transition, RNA binding involve dinposttranscriptional gene silencing, positive regulation of cell cycle G1-S phase transition, positive regulation of G1-S transition of mitotic cell cycle, trophoblast cell migration, T cell receptor complex, etc (Fig. 8C and Supplementary table S4). CBLL1 may participate in the progress of CHOL through methyl CPG binding, methylation dependent chromatin silencing, CIS trans isomerase activity, T cell receptor complex, positivere gulation of translational initiation, mRNA binding, etc (Fig. 8D and Supplementary table S5). CBLL1 may participate in the progress of DLBC through RNA binding involved in posttranscriptional gene silencing, negative regulation of tumor necrosis factor mediated signaling pathway, positive regulation of lipoprotein particle clearance, gene silencing, mRNA binding, positive regulation of cell aging, regulation of microglial cell activation, etc (Fig. 8F and Supplementary table S6). These results suggest that m6A related gene CBLL1 is strongly related to cancer.