# Single-cell RNA sequencing analysis of bone cancer pain model induced by Lewis lung cancer cells in male mice **Authors:** Qin Zhou, Jiahui Chen, Hui Wang, Ruyi Jin, Halisa Paerhati, Xiuxiu Zhu, Gaochao Dong, Minhao Zhang, Feng Jiang PMC · DOI: 10.1080/07853890.2026.2636340 · Annals of Medicine · 2026-02-27 ## TL;DR This study uses single-cell RNA sequencing to analyze spinal cord cell changes in a mouse model of bone cancer pain, revealing altered cell proportions and signaling pathways linked to pain and inflammation. ## Contribution The study provides novel insights into the cellular and molecular changes in the spinal cord associated with bone cancer pain using single-cell RNA sequencing. ## Key findings - BCP modeling increased microglia by 45% and oligodendrocytes by 43% in the L2-L4 spinal cord. - Significant changes in pain-related and inflammatory signaling pathways were observed in spinal cord cells. - Cell-cell interactions in the spinal cord were revealed, contributing to understanding BCP mechanisms. ## Abstract Bone cancer pain (BCP) is one of the most severe complications faced by cancer patients, with complex physiological and pathological mechanisms and unclear molecular characteristics. The BCP model was established by inoculating Lewis lung cancer cells into the femur to induce hyperalgesia and spontaneous pain. Single-cell RNA sequencing technology was used to characterize the cell composition and molecular features of the L2-L4 spinal cord after BCP modelling. Our research results identified a total of 10 cell types, namely excitatory neurons, inhibitory neurons, oligodendrocytes, oligodendrocyte precursor cells, Schwann cells, astrocytes, microglia, endothelial cells, fibroblasts, and pericytes. RNA sequencing analysis of the BCP model showed that the proportion of cells in the L2-L4 spinal cord changed significantly, with microglia increased by 45% and oligodendrocytes increased by 43%. Then, data were extracted from microglia, oligodendrocytes, blood-spinal cord barrier component cells (endothelial cells, pericytes, astrocytes), excitatory neurons, and inhibitory neurons, and differential genes were analysed and further enriched. The results suggest that the signalling pathways related to pain perception and transmission and promoting inflammation in the above cells have changed significantly. Finally, this study revealed the interaction between L2-L4 spinal cord cells in BCP. These data help to understand the molecular mechanism changes caused by BCP and contribute to the development of new treatment methods. ## Linked entities - **Diseases:** cancer (MONDO:0004992) - **Species:** Mus musculus (taxon 10090) ## Full-text entities - **Genes:** Adcy9 (adenylate cyclase 9) [NCBI Gene 11515] {aka AC9, ACtp10, D16Wsu65e, mKIAA0520}, Ptgds (prostaglandin D2 synthase (brain)) [NCBI Gene 19215] {aka 21kDa, L-PGDS, PGD2, PGDS, PGDS2, Ptgs3}, Apoe (apolipoprotein E) [NCBI Gene 25728] {aka APOEA}, B3galt2 (Beta-1,3-galactosyltransferase 2) [NCBI Gene 686081], Gpr160 (G protein-coupled receptor 160) [NCBI Gene 71862] {aka 1700025D19Rik, GPCR150}, CLDN5 (claudin 5) [NCBI Gene 7122] {aka AWAL, BEC1, CPETRL1, TMDVCF, TMVCF}, Pros1 (protein S (alpha)) [NCBI Gene 19128], Bcl2l1 (BCL2-like 1) [NCBI Gene 12048] {aka Bcl(X)L, Bcl-XL, Bcl2l, BclX, bcl-x, bcl2-L-1}, Tnfrsf21 (tumor necrosis factor receptor superfamily, member 21) [NCBI Gene 94185] {aka DR6, TR7}, TJP1 (tight junction protein 1) [NCBI Gene 7082] {aka ZO-1}, Degs1 (delta 4-desaturase, sphingolipid 1) [NCBI Gene 13244] {aka Degs, Des1, Mdes}, Phlpp1 (PH domain and leucine rich repeat protein phosphatase 1) [NCBI Gene 98432] {aka Phlpp, Plekhe1, SCOP, mKIAA0606}, P2ry12 (purinergic receptor P2Y, G-protein coupled 12) [NCBI Gene 70839] {aka 2900079B22Rik, 4921504D23Rik, P2Y12}, Lef1 (lymphoid enhancer binding factor 1) [NCBI Gene 16842] {aka 3000002B05, Lef-1}, Atp5po (ATP synthase peripheral stalk subunit OSCP) [NCBI Gene 28080] {aka ATPO, Atp5o, D12Wsu28e, OSCP}, Robo1 (roundabout guidance receptor 1) [NCBI Gene 19876] {aka DUTT1, Gm310}, Cacna1c (calcium channel, voltage-dependent, L type, alpha 1C subunit) [NCBI Gene 12288] {aka Cav1.2, Cchl1a1, D930026N18Rik, MBC, MELC-CC}, Grin2c (glutamate receptor, ionotropic, NMDA2C (epsilon 3)) [NCBI Gene 14813] {aka GluN2C, NMDAR2C, NR2C}, Gria1 (glutamate receptor, ionotropic, AMPA1 (alpha 1)) [NCBI Gene 14799] {aka 2900051M01Rik, Glr-1, Glr1, GluA1, GluR-A, GluRA}, Grin1 (glutamate receptor, ionotropic, NMDA1 (zeta 1)) [NCBI Gene 14810] {aka GluN1, GluRdelta1, GluRzeta1, M100174, NMD-R1, NMDAR1}, Magi2 (membrane associated guanylate kinase, WW and PDZ domain containing 2) [NCBI Gene 50791] {aka AIP-1, Acvri1, Acvrinp1, Acvrip1, Magi-2, S-SCAM}, Pik3cg (phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit gamma) [NCBI Gene 30955] {aka 5830428L06Rik, PI3Kgamma, p110gamma, p120-PI3K}, Angpt2 (angiopoietin 2) [NCBI Gene 11601] {aka Agpt2, Ang-2, Ang2}, Ryr2 (ryanodine receptor 2, cardiac) [NCBI Gene 20191] {aka 9330127I20Rik, RYR-2}, Cldn11 (claudin 11) [NCBI Gene 18417] {aka Claudin-11, Claudin11, Osp, Otm}, Il18 (interleukin 18) [NCBI Gene 29197] {aka IL-1 gamma, IL-18}, Cd14 (CD14 molecule) [NCBI Gene 60350], Pik3r1 (phosphoinositide-3-kinase regulatory subunit 1) [NCBI Gene 18708] {aka PI3K, p50alpha, p55alpha, p85alpha}, ANPEP (alanyl aminopeptidase, membrane) [NCBI Gene 290] {aka AP-M, AP-N, APN, CD13, GP150, LAP1}, Tfeb (transcription factor EB) [NCBI Gene 21425] {aka Tcfeb, bHLHe35}, Hspa5 (heat shock protein family A (Hsp70) member 5) [NCBI Gene 14828] {aka Bip, D2Wsu141e, D2Wsu17e, Grp78, Hsce70, SEZ-7}, Flt1 (FMS-like tyrosine kinase 1) [NCBI Gene 14254] {aka Flt-1, VEGFR-1, VEGFR1, sFlt1}, Tnfrsf19 (tumor necrosis factor receptor superfamily, member 19) [NCBI Gene 29820] {aka TAJ, TAJ-ALPHA, TRADE, Troy}, Cldn5 (claudin 5) [NCBI Gene 12741] {aka MBEC1, Tmvcf}, F11r (F11 receptor) [NCBI Gene 16456] {aka 9130004G24, ESTM33, JAM, JAM-1, JAM-A, Jcam}, Nrxn1 (neurexin I) [NCBI Gene 18189] {aka 1700062G21Rik, 9330127H16Rik, A230068P09Rik, mKIAA0578}, Tac2 (tachykinin 2) [NCBI Gene 21334] {aka PPT-B, Tac3}, Ncam1 (neural cell adhesion molecule 1) [NCBI Gene 17967] {aka CD56, E-NCAM, NCAM-1, Ncam}, Slit2 (slit guidance ligand 2) [NCBI Gene 20563] {aka Drad-1, E030015M03Rik, E130320P19Rik, Slil3, b2b1200.1Clo, mKIAA4141}, Scd2 (stearoyl-Coenzyme A desaturase 2) [NCBI Gene 20250] {aka Mir5114, Scd-2, mir-5114, swty}, Mag (myelin-associated glycoprotein) [NCBI Gene 17136] {aka Gma, siglec-4a}, Plcb4 (phospholipase C, beta 4) [NCBI Gene 18798] {aka A930039J07Rik, C230058B11Rik}, Adipor2 (adiponectin receptor 2) [NCBI Gene 68465] {aka 1110001I14Rik, ADCR2, D6Ucla1e, Paqr2}, Vtn (vitronectin) [NCBI Gene 22370] {aka Vn}, Tyrobp (TYRO protein tyrosine kinase binding protein) [NCBI Gene 22177] {aka DAP12, KARAP, Ly83}, Gper1 (G protein-coupled estrogen receptor 1) [NCBI Gene 76854] {aka 6330420K13Rik, CMKRL2, Ceprl, FEG-1, GPCR-Br, Gper}, Prkacb (protein kinase, cAMP dependent, catalytic, beta) [NCBI Gene 18749] {aka CbPKA, PKA C-beta, Pkacb}, Ackr3 (atypical chemokine receptor 3) [NCBI Gene 12778] {aka CXC-R7, CXCR-7, Cmkor1, Cxcr7, RDC-1, Rdc1}, Cd81 (CD81 antigen) [NCBI Gene 12520] {aka Tapa-1, Tapa1, Tspan28}, Map3k4 (mitogen-activated protein kinase kinase kinase 4) [NCBI Gene 26407] {aka D17Rp17, D17Rp17e, MAPKKK4, MEKK 4, MTK1, Mek4b}, Ptprm (protein tyrosine phosphatase receptor type M) [NCBI Gene 19274] {aka RPTPmu, mKIAA4044}, Gapdh (glyceraldehyde-3-phosphate dehydrogenase) [NCBI Gene 14433] {aka Gapd}, Hexb (hexosaminidase subunit beta) [NCBI Gene 294673], Akt1 (Akt serine/threonine kinase 1) [NCBI Gene 11651] {aka Akt, LTR-akt, PKB, PKB/Akt, PKBalpha, Rac}, Nlk (nemo like kinase) [NCBI Gene 18099], Ctnna3 (catenin alpha 3) [NCBI Gene 216033] {aka 4930429L08Rik, 4933408A16, Catna3, Vr22}, Il10 (interleukin 10) [NCBI Gene 25325] {aka IL10X, If2a}, Itpr1 (inositol 1,4,5-trisphosphate receptor 1) [NCBI Gene 16438] {aka D6Pas2, Gm10429, IP3R 1, IP3R1, InsP3R, Ip3r}, Apln (apelin) [NCBI Gene 30878] {aka 6030430G11Rik, Apel}, Rac1 (Rac family small GTPase 1) [NCBI Gene 19353] {aka D5Ertd559e} - **Diseases:** ALS (MESH:D008113), metabolic disorder (MESH:D008659), multiple sclerosis (MESH:D009103), SCI (MESH:D013119), heat hyperalgesia (MESH:D006930), acne (MESH:D000152), spine hyperplasia (MESH:D006965), cancer pain (MESH:D000072716), Spinal cord neuron dysfunction (MESH:D013118), Lewis lung carcinoma (MESH:D018827), dysfunction (MESH:D006331), hypersensitivity (MESH:D004342), neuronal damage (MESH:D009410), disorder of oligodendrocytes (MESH:D056784), neuropathic pain (MESH:D009437), bone destruction (MESH:D001847), NOD (MESH:D020191), neuroinflammation (MESH:D000090862), AD (MESH:D000544), metabolic failure (MESH:D051437), diabetic neuropathy (MESH:D003929), neurotoxicity (MESH:D020258), Lewis lung cancer (MESH:D008175), diabetes (MESH:D003920), Tumour (MESH:D009369), mechanical abnormal pain (MESH:D010146), myelin dysfunction (MESH:D003711), lipid metabolism (MESH:D052439), BCP (MESH:D001859), Parkinson's (MESH:D010300), tremors (MESH:D014202), mitochondrial (MESH:D028361), degenerative diseases (MESH:D019636), inflammation (MESH:D007249), bone metastasis (MESH:D009362) - **Chemicals:** glucose (MESH:D005947), ethanol (MESH:D000431), formalin (MESH:D005557), calcium (MESH:D002118), GABA (MESH:D005680), eosin (MESH:D004801), sphingolipid (MESH:D013107), alcohol (MESH:D000438), glutamate (MESH:D018698), paraformaldehyde (MESH:C003043), lipid (MESH:D008055), isoflurane (MESH:D007530), ATP (MESH:D000255), Evans blue (MESH:D005070), CO2 (MESH:D002245), streptomycin (MESH:D013307), EDTA (MESH:D004492), penicillin (MESH:D010406), Haematoxylin (MESH:D006416), BSCB (-), iodophor (MESH:D007466), paraffin (MESH:D010232), H&E (MESH:D006371) - **Species:** Homo sapiens (human, species) [taxon 9606], Rattus norvegicus (brown rat, species) [taxon 10116], Mus musculus (house mouse, species) [taxon 10090] - **Mutations:** A 30 G - **Cell lines:** LLC-Luc — Mus musculus (Mouse), Malignant tumors of the mouse pulmonary system, Cancer cell line (CVCL_5653), /6J — Homo sapiens (Human), Cutaneous melanoma, Cancer cell line (CVCL_W797), Lewis lung cancer — Mus musculus (Mouse), Malignant tumors of the mouse pulmonary system, Cancer cell line (CVCL_4358) ## Full text _Full body text omitted from this summary view._ Fetch the complete paper as Markdown: https://tomesphere.com/paper/PMC12951664/full.md ## Figures 10 figures with captions in the complete paper: https://tomesphere.com/paper/PMC12951664/full.md ## References 56 references — full list in the complete paper: https://tomesphere.com/paper/PMC12951664/full.md --- Source: https://tomesphere.com/paper/PMC12951664