犬乳腺肿瘤细胞CHMM和CHMP的差异表达蛋白筛选

doi:10.16431/j.cnki.1671-7236.2026.01.043

Abstract

Abstract:

Objective This study aimed to screen differentially expressed proteins between canine mammary tumor cells CHMM and CHMP， so as to provide scientific basis for the search of effective biomarkers for the diagnosis and treatment of canine mammary tumors. Method Using canine mammary tumor thoracic metastatic cells CHMM and primary cells CHMP as research objects， the morphologies， cloning ability， and migration ability of the two cell lines were compared through microscopic observation， cloning experiment， and Transwell experiment. Then， differentially expressed proteins in cells were screened using high-performance liquid chromatography-mass spectrometry analysis technology. GO function and KEGG pathway enrichment analysis on differentially expressed proteins was performed. Protein-protein interaction analysis was performed on differentially expressed proteins using STRING database. The proteomics results were validated through Western blotting experiment. Result CHMP cells exhibited a long spindle shape with single-cell growth， while CHMM cells grew in clusters with relatively small cells. CHMP cells had stronger migration ability， while CHMM cells had stronger cloning ability. A total of 2 049 proteins with differential expression were screened by sequencing （FoldChange>1.2 and P<0.05）， including 1 161 down regulated proteins and 888 upregulated proteins. GO functional enrichment analysis results showed that the differentially expressed proteins in cells involvement in biological processes mainly included cellular process， single-organism process， metabolic process，regulation of biological process， response to stimulus， etc. The cellular components included cell， cell part， organelle， organelle part， etc. Molecular functions mainly included catalytic activity， binding， and transporter activity. KEGG pathway enrichment analysis showed that the differentially expressed proteins in cells mainly participated in metabolic pathway signaling pathways. Differentially expressed proteins interaction analysis showed that CD44， MMP14 and DPYSL3 proteins had an interaction relationship. Western blotting showed that compared with CHMM cells， the expression level of GSDMD in CHMP cells significantly increased and the expression level of CD44 significantly decreased （P<0.05）， which was consistent with proteomic results，confirming the accuracy of sequencing. Conclusion Through proteomics screening， 1 161 downregulated proteins such as GSDMD and 888 upregulated proteins such as CD44 were identified in canine mammary tumor cells CHMM and CHMP， involving multiple biological processes such as metabolic pathways. The results of this study could provide scientific basis for the diagnosis and treatment of canine mammary tumors.

Key words: canine mammary gland tumor cells; proteomics; differentially expressed protein; metabolic pathways

CLC Number:

S857.4

YU Zhiying, SU Chunyang, XU Enshuang, ZHENG Jiasan. Differentially Expressed Protein Screening of Canine Mammary Tumor Cells CHMM and CHMP[J]. China Animal Husbandry & Veterinary Medicine, 2026, 53(1): 479-488.

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References 20

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蛋白名称 Protein name	P值 P-value	模式 Pattern	蛋白名称 Protein name	P值 P-value	模式 Pattern
蛋白磷酸酶1调节亚基14C PPP1R14C	2.20E-16	上调	细胞黏附因子 CD44	5.86E-15	下调
跨膜单链糖蛋白 TF	4.87E-11	上调	高密度脂蛋白结合蛋白 HDLBP	8.64E-15	下调
Gasdermin D蛋白 GSDMD	5.10E-11	上调	平滑肌蛋白22 TAGLN	1.10E-14	下调
L抗原家族成员 LAGE3	6.16E-11	上调	二氢嘧啶酶样 3 DPYSL3	1.20E-14	下调
纤毛和鞭毛相关蛋白20 CFAP20	9.17E-11	上调	蛋白质S100-A4 S100A4	2.72E-14	下调
SUMO特异性肽酶3 SENP3	1.71E-10	上调	侵袭基质金属蛋白酶14 MMP14	3.85E-14	下调
丝氨酸B10 SERPINB10	2.30E-10	上调	α-辅肌动蛋白-1 ACTN1	4.90E-14	下调
人细胞角蛋白 Keratin 14	6.31E-10	上调	胸腺肽β-4 TMSB4	4.05E-13	下调
碳酰还原酶 LOC610164	4.31E-10	上调	OCIA域2 OCIAD2	4.59E-13	下调
细胞分裂周期蛋白 73 CDC73	7.36E-10	上调	FCH和双SH3结构域1 FCHSD1	5.35E-13	下调

通路 Pathways	蛋白数量 Number of proteins	P值 P-value	通路ID Pathway ID
代谢途径Metabolic pathways	629（12.66%）	0.9857873	ko01100
内吞作用Endocytosis	186（3.74%）	0.8479323	ko04144
癌症通路Pathways in cancer	165（3.32%）	0.01051816	ko05200
RNA转运RNA transport	135（2.72%）	1	ko03013
内质网中的蛋白质加工 Protein processing in endoplasmic reticulum	131（2.64%）	0.8149936	ko04141
剪接体Spliceosome	129（2.60%）	0.9999984	ko03040
亨廷顿氏疾病 Huntington’s disease	126（2.54%）	0.9885915	ko05016
人类疱疹病毒4型感染 Epstein-Barr virus infection	122（2.46%）	0.9238041	ko05169
核糖体Ribosome	122（2.46%）	0.9954844	ko03010
肌动蛋白细胞骨架调控 Regulation of actin cytoskeleton	121（2.44%）	0.03166765	ko04810
PI3K-Akt信号通路 PI3K-Akt signaling pathway	115（2.31%）	0.2105181	ko04151
细胞黏附Focal adhesion	113（2.27%）	0.00006281731	ko04510

Differentially Expressed Protein Screening of Canine Mammary Tumor Cells CHMM and CHMP

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