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STAT3-coordinated migration facilitates the dissemination of diffuse large B-cell lymphomas

Pan, Yi-Ru, Chen, Chih-Cheng, Chan, Yu-Tien, Wang, Hsiao-Jung, Chien, Fan-Tso, Chen, Yeng-Long, Liu, Jing-Lan, Yang, Muh-Hwa·2018·Nature Communications
DOI 10.1038/s41467-018-06134-zPMC PMC6135800
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Abstract

The motile characteristics and mechanisms that drive the dissemination of diffuse large B-cell lymphoma (DLBCL) are elusive. Here, we show that DLBCL initiates dissemination through activating STAT3-mediated amoeboid migration. Mechanistically, STAT3 activates RHOH transcription, which competes with the RhoGDP dissociation inhibitor RhoGDIγ to activate RhoA. In addition, activated STAT3 regulates microtubule dynamics and releases ARHGEF2 to activate RhoA. Both the JAK inhibitor ruxolitinib and the microtubule stabilizer Taxol suppress DLBCL cell dissemination in vivo. A clinical DLBCL sample analysis shows that STAT3-driven amoeboid movement is particularly important for the transition from stage I to stage II. This study elucidates the mechanism of DLBCL dissemination and progression and highlights the potential of combating advanced DLBCL with a JAK/STAT inhibitor or microtubule stabilizer to reduce DLBCL motility; these findings may have a great impact on the development of patient-tailored treatments for DLBCL.

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Fig. 1 Amoeboid movement is involved in the early dissemination of DLBCL. a GSEA for examining the association between the amoeboid movement signature and gene expression array from DLBCL patients with stage II–IV vs. stage I (GSE11318). b Distribution of the H score for MLC S19 phosphorylation (MLCpS19) from DLBCL patients. n = 72. c Representative results of immunohistochemical staining for MLC S19 phosphorylation. Scale bar = 20 μm. d SU-DHL-5, OCI-Ly3, HT, U2932, and DB cells were grown in collagen gels and OEC-M1 cells were grown on top of collagen gels overnight. Upper and middle: representative phase-contrast images. Scale bar = 50 μm (Upper) or 10 μm (middle). Lower: immunofluorescent staining for F-actin (red). Blue, nuclei. Scale bar = 5 μm. e Quantification of movement speeds of lymphoma cell lines treated with inhibitors (n = 20). PI: proteinase inhibitor. f Quantification of movement speeds of OCI-Ly3 cells treated with inhibitors (n = 20). *P < 0.05, **P < 0.005 by Student’s t-test; Kolmogorov–Smirnov tests for GSEA. See also Supplementary Fig. 1 and Supplementary Movie 1. See Supplementary Table 1 for patient demographics in b and c; Supplementary Data 2 for amoeboid signature in a; Supplementary Table 4 for working concentration of inhibitors in e and f
Fig. 1 Amoeboid movement is involved in the early dissemination of DLBCL. a GSEA for examining the association between the amoeboid movement signature and gene expression array from DLBCL patients with stage II–IV vs. stage I (GSE11318). b Distribution of the H score for MLC S19 phosphorylation (MLCpS19) from DLBCL patients. n = 72. c Representative results of immunohistochemical staining for MLC S19 phosphorylation. Scale bar = 20 μm. d SU-DHL-5, OCI-Ly3, HT, U2932, and DB cells were grown in collagen gels and OEC-M1 cells were grown on top of collagen gels overnight. Upper and middle: representative phase-contrast images. Scale bar = 50 μm (Upper) or 10 μm (middle). Lower: immunofluorescent staining for F-actin (red). Blue, nuclei. Scale bar = 5 μm. e Quantification of movement speeds of lymphoma cell lines treated with inhibitors (n = 20). PI: proteinase inhibitor. f Quantification of movement speeds of OCI-Ly3 cells treated with inhibitors (n = 20). *P < 0.05, **P < 0.005 by Student’s t-test; Kolmogorov–Smirnov tests for GSEA. See also Supplementary Fig. 1 and Supplementary Movie 1. See Supplementary Table 1 for patient demographics in b and c; Supplementary Data 2 for amoeboid signature in a; Supplementary Table 4 for working concentration of inhibitors in e and fCC BY · PubMed Central
Fig. 2 Amoeboid movement mediates DLBCL dissemination in vivo. a A schema for showing the establishment of DLBCL sublines with increased tumorigenic and disseminating capability. b Mice experiments for generating the HT sublines. HT-1 subline cells were injected into the splenic parenchyma, and the bioluminescent signals were detected on day 56. c Quantification of movement speeds of different sublines (n = 20). d Quantification of movement speeds of different sublines treated with corresponding inhibitors (n = 20). e Investigation of the in vivo dissemination of the HT-S1 or HT-L1 sublines. Cells were injected into the splenic parenchyma. Mice bioluminescent signals on indicated days were detected (n = 6; other 4 mice in Supplementary Fig. 2d). f Quantification of disseminated numbers (red dotted lines) in e and Supplementary Fig. 2d. g Investigation of the in vivo dissemination of HT-L1 receiving shRNAs specific to ROCK I (shROCK; clones #1, #2) or a scramble sequence (shScr). The cells were injected into the splenic parenchyma. Mice bioluminescent signals on indicated days were detected (n = 6; other 4 mice in Supplementary Fig. 2h). h Quantification of the numbers disseminated lesions in g and Supplementary Fig. 2h. i, j Quantification of bioluminescent intensity of g and Supplementary Fig. 2h from the primary sites (yellow dotted line circled area in photos)(i) and the disseminated sites (red dotted lines circled area in photos) (j). NS, not significant. k Quantification of the CTCs (circulating tumor cells) collected from the mice described in panel g (n = 6). *P < 0.05, **P < 0.005 by Student’s t-test. See also Supplementary Fig. 2 and Supplementary Movie 2. See Supplementary Table 4 for working concentration of inhibitors
Fig. 2 Amoeboid movement mediates DLBCL dissemination in vivo. a A schema for showing the establishment of DLBCL sublines with increased tumorigenic and disseminating capability. b Mice experiments for generating the HT sublines. HT-1 subline cells were injected into the splenic parenchyma, and the bioluminescent signals were detected on day 56. c Quantification of movement speeds of different sublines (n = 20). d Quantification of movement speeds of different sublines treated with corresponding inhibitors (n = 20). e Investigation of the in vivo dissemination of the HT-S1 or HT-L1 sublines. Cells were injected into the splenic parenchyma. Mice bioluminescent signals on indicated days were detected (n = 6; other 4 mice in Supplementary Fig. 2d). f Quantification of disseminated numbers (red dotted lines) in e and Supplementary Fig. 2d. g Investigation of the in vivo dissemination of HT-L1 receiving shRNAs specific to ROCK I (shROCK; clones #1, #2) or a scramble sequence (shScr). The cells were injected into the splenic parenchyma. Mice bioluminescent signals on indicated days were detected (n = 6; other 4 mice in Supplementary Fig. 2h). h Quantification of the numbers disseminated lesions in g and Supplementary Fig. 2h. i, j Quantification of bioluminescent intensity of g and Supplementary Fig. 2h from the primary sites (yellow dotted line circled area in photos)(i) and the disseminated sites (red dotted lines circled area in photos) (j). NS, not significant. k Quantification of the CTCs (circulating tumor cells) collected from the mice described in panel g (n = 6). *P < 0.05, **P < 0.005 by Student’s t-test. See also Supplementary Fig. 2 and Supplementary Movie 2. See Supplementary Table 4 for working concentration of inhibitorsCC BY · PubMed Central
Fig. 3 JAK-STAT3 signaling is the major pathway mediating cell movement in DLBCL. a A schema for showing the mining process for identification of the signaling pathways that regulate amoeboid movement in DLBCL. See Supplementary Table 3 for the information and references for the signal pathways. b A heat map showing the relative cell movement of SU-DHL-5 cells upon the treatment of inhibitors. The velocity of the cells was measured and expressed as fold relative the level control cells. Red, decreased motility level; blue, increased motility level. c A GSEA plot for showing the association between STAT3-regulated signature (Supplementary Data 2) and DLBCL stage I vs. II–IV (GSE11318). d Western blots for showing the efficacy of OCI-Ly3 cells treated with indicated inhibitors. e Quantification of movement speeds of inhibitor treated OCI-Ly3 cells (n = 20). f Western blots for showing the knockdown efficacy of HT-L1 and HT-L2 sublines receiving shRNAs specific to STAT3 (shSTAT3) or scramble (shScr). g Quantification of movement speeds of the cells described in panel f (n = 30). h Western blots for showing the transfection efficacy of HT cells expressing FLAG-STAT3 WT and its mutants. i Quantification of movement speeds of cells described in panel h (n = 20). j Quantification of movement speeds of HT cells expressing STAT3 WT, Y640F or control vector (puro) treated H1152 or DMSO (n = 30). k–l Representative results of western blots and pull down assay for showing the level of Y705-phosphorylated STAT3, STAT3, α-tubulin, active RhoA, and total RhoA in SU-DHL-5 cells under the treatment of different inhibitors for 6 h (k). Quantification of RhoA activity from three independent experiments (l). Data represent mean ± S.D. m Western blots and pull down assay for showing the level of STAT3, α-tubulin, active RhoA, and total RhoA in SU-DHL-5 cells receiving shRNA against STAT3 or a scramble sequence. *P < 0.05, **P < 0.005 by Student’s t-test; Kolmogorov–Smirnov tests for GSEA. See also Supplementary Fig. 3 and Supplementary Movie 3. See Supplementary Table 4 for working concentration of inhibitors
Fig. 3 JAK-STAT3 signaling is the major pathway mediating cell movement in DLBCL. a A schema for showing the mining process for identification of the signaling pathways that regulate amoeboid movement in DLBCL. See Supplementary Table 3 for the information and references for the signal pathways. b A heat map showing the relative cell movement of SU-DHL-5 cells upon the treatment of inhibitors. The velocity of the cells was measured and expressed as fold relative the level control cells. Red, decreased motility level; blue, increased motility level. c A GSEA plot for showing the association between STAT3-regulated signature (Supplementary Data 2) and DLBCL stage I vs. II–IV (GSE11318). d Western blots for showing the efficacy of OCI-Ly3 cells treated with indicated inhibitors. e Quantification of movement speeds of inhibitor treated OCI-Ly3 cells (n = 20). f Western blots for showing the knockdown efficacy of HT-L1 and HT-L2 sublines receiving shRNAs specific to STAT3 (shSTAT3) or scramble (shScr). g Quantification of movement speeds of the cells described in panel f (n = 30). h Western blots for showing the transfection efficacy of HT cells expressing FLAG-STAT3 WT and its mutants. i Quantification of movement speeds of cells described in panel h (n = 20). j Quantification of movement speeds of HT cells expressing STAT3 WT, Y640F or control vector (puro) treated H1152 or DMSO (n = 30). k–l Representative results of western blots and pull down assay for showing the level of Y705-phosphorylated STAT3, STAT3, α-tubulin, active RhoA, and total RhoA in SU-DHL-5 cells under the treatment of different inhibitors for 6 h (k). Quantification of RhoA activity from three independent experiments (l). Data represent mean ± S.D. m Western blots and pull down assay for showing the level of STAT3, α-tubulin, active RhoA, and total RhoA in SU-DHL-5 cells receiving shRNA against STAT3 or a scramble sequence. *P < 0.05, **P < 0.005 by Student’s t-test; Kolmogorov–Smirnov tests for GSEA. See also Supplementary Fig. 3 and Supplementary Movie 3. See Supplementary Table 4 for working concentration of inhibitorsCC BY · PubMed Central
Fig. 4 The IL10-JAK-STAT3 positive feedback loop contributes to DLBCL motility. a, b Representative results of western blots of indicated proteins in whole-cell lysates (WCL), IL-6 and IL-10 in conditioned media (CM) from HT-S1 and HT-L1 sublines. Quantification of IL-6 and IL-10 expression. n = 3. Data represent mean ± S.D. c, d OCI-Ly3 cells were grown in collagen gels and treated with IL-6 or IL-10. Representative western blots and pull down assay for showing the level of indicated proteins. Quantification of RhoA activity. n = 3. Data represent mean ± S.D. e Quantification of movement speeds of OCI-Ly3 cells treated with IL-6 or IL-10 (n = 30). f Quantification of movement speeds of OCI-Ly3 cells treated with IgG, IL-6, or IL-10 neutralizing antibody (n = 30). g GSEA for showing the association between the signature of amoeboid movement and gene expression array from healthy peripheral blood mononuclear cells stimulated with IL-10 or control media (GSE43700). h GSEA for showing the association between the signature of amoeboid movement and gene expression array from monocyte-derived macrophages stimulated with IL-6 or control media (GSE8515). i HT cells expressing WT STAT3 or a control vector (puro) were injected into the spleen of SCID/beige mice. Mice bioluminescent signals on indicated days were detected (n = 8; other 5 mice in Supplementary Fig. 4i). j Quantification of disseminated numbers in i and Supplementary Fig. 4i. k-l Quantification of bioluminescent intensity of i and Supplementary Fig. 4i. from the primary sites (yellow dotted lines circled area) and the disseminated sites (red dotted lines circled area). NS, not significant. m Quantification of the circulating tumor cells (CTCs) collected from the mice described in panel i. n = 8. n HT cells expressing WT STAT3 or a control vector (puro) were intraperitoneal injected into SCID/beige mice for 9 weeks. The dot plot shows the serum concentrations of IL-10. n = 6. o Distribution of the H score for IL-10 expression from 73 DLBCL patients. p Representative results of immunohistochemical staining for IL-10. Scale bar = 20 μm.*, P < 0.05; **, P < 0.005 by Student’s t-test; Kolmogorov–Smirnov tests for GSEA. See also Supplementary Fig. 4
Fig. 4 The IL10-JAK-STAT3 positive feedback loop contributes to DLBCL motility. a, b Representative results of western blots of indicated proteins in whole-cell lysates (WCL), IL-6 and IL-10 in conditioned media (CM) from HT-S1 and HT-L1 sublines. Quantification of IL-6 and IL-10 expression. n = 3. Data represent mean ± S.D. c, d OCI-Ly3 cells were grown in collagen gels and treated with IL-6 or IL-10. Representative western blots and pull down assay for showing the level of indicated proteins. Quantification of RhoA activity. n = 3. Data represent mean ± S.D. e Quantification of movement speeds of OCI-Ly3 cells treated with IL-6 or IL-10 (n = 30). f Quantification of movement speeds of OCI-Ly3 cells treated with IgG, IL-6, or IL-10 neutralizing antibody (n = 30). g GSEA for showing the association between the signature of amoeboid movement and gene expression array from healthy peripheral blood mononuclear cells stimulated with IL-10 or control media (GSE43700). h GSEA for showing the association between the signature of amoeboid movement and gene expression array from monocyte-derived macrophages stimulated with IL-6 or control media (GSE8515). i HT cells expressing WT STAT3 or a control vector (puro) were injected into the spleen of SCID/beige mice. Mice bioluminescent signals on indicated days were detected (n = 8; other 5 mice in Supplementary Fig. 4i). j Quantification of disseminated numbers in i and Supplementary Fig. 4i. k-l Quantification of bioluminescent intensity of i and Supplementary Fig. 4i. from the primary sites (yellow dotted lines circled area) and the disseminated sites (red dotted lines circled area). NS, not significant. m Quantification of the circulating tumor cells (CTCs) collected from the mice described in panel i. n = 8. n HT cells expressing WT STAT3 or a control vector (puro) were intraperitoneal injected into SCID/beige mice for 9 weeks. The dot plot shows the serum concentrations of IL-10. n = 6. o Distribution of the H score for IL-10 expression from 73 DLBCL patients. p Representative results of immunohistochemical staining for IL-10. Scale bar = 20 μm.*, P < 0.05; **, P < 0.005 by Student’s t-test; Kolmogorov–Smirnov tests for GSEA. See also Supplementary Fig. 4CC BY · PubMed Central
Fig. 5 STAT3 induces RHOH transcription to activate RhoA through competitive interaction with RhoGDIγ. a Western blots of RhoH in SU-DHL-5 cells receiving shRNAs specific to RhoH (shRhoH; clones #1, #2) or a scramble sequence (shScr). b Quantification of movement speeds (n = 30). c, d Western blots for the indicated proteins and pull down assay for GTP-bound RhoA in SU-DHL-5 cells receiving shRhoH or shScr (c). Quantification of RhoA activity (n = 3) (d). e, f Pull down assay showing the GTP-RhoA in HT cells transfected with HA-RhoH or a control vector (e). Quantification of RhoA activity (n = 3) (f). g, h Analysis of the mRNA (g) (n = 7) and protein (h) expression of STAT3 and RhoH in SU-DHL-5 cells receiving shRNAs against STAT3 or a scramble sequence. i, j Analysis of the mRNA (i) (n = 4) and protein (j) expression of STAT3 and RhoH in HT cells stably expressed FLAG-STAT3 WT or control vector. k Western blots for showing the efficacy of inhibitors and the expression level of RhoH in SU-DHL-5 cells expressing HA-RhoH or control vector. l Quantification of the movement speeds (n = 20). m Luciferase reporter assay. The result is presented as luciferase activity/β-galactosidase (β-gal). n = 4. A schema for the promoter region of the RHOH gene and reporter constructs. *mutated sites. n ChIP assay in SU-DHL-5 after ruxolitinib treatment for 24 h. Primer 1, control. Primer 2, amplifying fragment containing STAT3-binding element (−992–−983, TTCTTTGAA). n = 4. o Quantification of movement speeds of SU-DHL-5 cells receiving shRNAs specific to RhoGDIγ (shRhoGDI-γ; clones #1, #2) or shScr. n = 30. p, q Western blots and the pull down assay for GTP-bound RhoA in HA-RhoGDI-γ or control vector transfected HT cells (p). Quantification of RhoA activity (n = 3) (q). r Immunoprecipitation-western blot of HT cells stably expressed FLAG-RhoA transfected with indicated plasmids. s Western blots of RhoGDIγ and RhoH in HT cells receiving shRNAs against RhoH, RhoGDIγ, or a scramble sequence. t Quantification of movement speeds (n = 30). Data represent mean ± S.D. in panels d, f, g, i, m, n, and q. *P < 0.05, **P < 0.005 by Student’s t-test. See also Supplementary Fig. 5 and Supplementary Movie 4
Fig. 5 STAT3 induces RHOH transcription to activate RhoA through competitive interaction with RhoGDIγ. a Western blots of RhoH in SU-DHL-5 cells receiving shRNAs specific to RhoH (shRhoH; clones #1, #2) or a scramble sequence (shScr). b Quantification of movement speeds (n = 30). c, d Western blots for the indicated proteins and pull down assay for GTP-bound RhoA in SU-DHL-5 cells receiving shRhoH or shScr (c). Quantification of RhoA activity (n = 3) (d). e, f Pull down assay showing the GTP-RhoA in HT cells transfected with HA-RhoH or a control vector (e). Quantification of RhoA activity (n = 3) (f). g, h Analysis of the mRNA (g) (n = 7) and protein (h) expression of STAT3 and RhoH in SU-DHL-5 cells receiving shRNAs against STAT3 or a scramble sequence. i, j Analysis of the mRNA (i) (n = 4) and protein (j) expression of STAT3 and RhoH in HT cells stably expressed FLAG-STAT3 WT or control vector. k Western blots for showing the efficacy of inhibitors and the expression level of RhoH in SU-DHL-5 cells expressing HA-RhoH or control vector. l Quantification of the movement speeds (n = 20). m Luciferase reporter assay. The result is presented as luciferase activity/β-galactosidase (β-gal). n = 4. A schema for the promoter region of the RHOH gene and reporter constructs. *mutated sites. n ChIP assay in SU-DHL-5 after ruxolitinib treatment for 24 h. Primer 1, control. Primer 2, amplifying fragment containing STAT3-binding element (−992–−983, TTCTTTGAA). n = 4. o Quantification of movement speeds of SU-DHL-5 cells receiving shRNAs specific to RhoGDIγ (shRhoGDI-γ; clones #1, #2) or shScr. n = 30. p, q Western blots and the pull down assay for GTP-bound RhoA in HA-RhoGDI-γ or control vector transfected HT cells (p). Quantification of RhoA activity (n = 3) (q). r Immunoprecipitation-western blot of HT cells stably expressed FLAG-RhoA transfected with indicated plasmids. s Western blots of RhoGDIγ and RhoH in HT cells receiving shRNAs against RhoH, RhoGDIγ, or a scramble sequence. t Quantification of movement speeds (n = 30). Data represent mean ± S.D. in panels d, f, g, i, m, n, and q. *P < 0.05, **P < 0.005 by Student’s t-test. See also Supplementary Fig. 5 and Supplementary Movie 4CC BY · PubMed Central
Fig. 6 STAT3 activates RhoA through regulating microtubule dynamics. a, c Western blots for showing the level of Y705-phosphorylated STAT3, total STAT3, and acetylated tubulin in HT-L1 vs. HT-S1 (a) and HT-L2 vs. HT-S2 (c) sublines. b, d Quantification of relative level of acetylated tubulin in a and c, respectively (n = 3). e, f Fluorescence recovery after photobleaching (FRAP) of STAT3 WT-expressed HT cells stably expressing cherry-tubulin. The inhibitors were added 4 h before FRAP (e). FRAP of HT-STAT3 cells, IL-10-treated HT-STAT3 cells, and IL-10-treated HT-STAT3 cells with S31-201 which stably expressed cherry-tubulin. The cells were grown in collagen gels in serum-free medium and then treated with IL-10 (f). Quantification of fluorescence intensity before and after bleaching. The graph illustrates fluorescence recovery curves. (n ≥ 25). g Quantification of movement speeds of SU-DHL-5, OCI-Ly3, or HT cells treated with different inhibitors (n ≥ 20). h Quantification of movement speeds of SU-DHL-5 cells treated with different inhibitors (n = 20). i, l Representative results of western blots and pull down assay for showing the level of Y705-phosphorylated STAT3, acetylated tubulin, and active (GTP-bound) Rho family proteins in OCI-Ly3 cells under the treatment of different inhibitors for 6 h. j, m Quantification of relative level of acetylated tubulin in i and l, respectively (n = 3). k, n Quantification of RhoA activity in i and l, respectively (n = 3). Data represent mean ± S.D. in panels b, d, e, f, j, k, m, and n. *P < 0.05, **P < 0.005 by Student’s t-test. See also Supplementary Fig. 6 and Supplementary Movie 5. See Supplementary Table 4 for the working concentrations of inhibitors
Fig. 6 STAT3 activates RhoA through regulating microtubule dynamics. a, c Western blots for showing the level of Y705-phosphorylated STAT3, total STAT3, and acetylated tubulin in HT-L1 vs. HT-S1 (a) and HT-L2 vs. HT-S2 (c) sublines. b, d Quantification of relative level of acetylated tubulin in a and c, respectively (n = 3). e, f Fluorescence recovery after photobleaching (FRAP) of STAT3 WT-expressed HT cells stably expressing cherry-tubulin. The inhibitors were added 4 h before FRAP (e). FRAP of HT-STAT3 cells, IL-10-treated HT-STAT3 cells, and IL-10-treated HT-STAT3 cells with S31-201 which stably expressed cherry-tubulin. The cells were grown in collagen gels in serum-free medium and then treated with IL-10 (f). Quantification of fluorescence intensity before and after bleaching. The graph illustrates fluorescence recovery curves. (n ≥ 25). g Quantification of movement speeds of SU-DHL-5, OCI-Ly3, or HT cells treated with different inhibitors (n ≥ 20). h Quantification of movement speeds of SU-DHL-5 cells treated with different inhibitors (n = 20). i, l Representative results of western blots and pull down assay for showing the level of Y705-phosphorylated STAT3, acetylated tubulin, and active (GTP-bound) Rho family proteins in OCI-Ly3 cells under the treatment of different inhibitors for 6 h. j, m Quantification of relative level of acetylated tubulin in i and l, respectively (n = 3). k, n Quantification of RhoA activity in i and l, respectively (n = 3). Data represent mean ± S.D. in panels b, d, e, f, j, k, m, and n. *P < 0.05, **P < 0.005 by Student’s t-test. See also Supplementary Fig. 6 and Supplementary Movie 5. See Supplementary Table 4 for the working concentrations of inhibitorsCC BY · PubMed Central
Fig. 7 Validation of the JAK/STAT3-microtubule-RhoA axis in DLBCL in vivo and clinical samples. a A schema for showing the experimental design of the mice experiment. The HT-1 subline was injected into the spleen of SCID/beige mice. Before Taxol or Ruxolitinib injection, the primary tumors were removed. The drugs were injected intraperitoneally on Day 0 and bioluminescent signals were taken at the indicated time points. NaCl, vehicle control. n = 6 for each group. b Quantification of disseminated lesions (red dotted lines) in a. c Representative pictures of immunohistochemical staining of the Y705-phosphorylated STAT3, and S19-phosphorylated MCL, RhoH, and CD20 in DLBCL cases. The clinical stage of the representative cases is indicated. Scale bar = 25 μm. d Distribution of H scores of Y705-phosphorylated STAT3 in DLBCL patients with different stages. e Distribution of H scores of RhoH in DLBCL patients with different stages. *P < 0.05 by Student’s t-test. See also Supplementary Fig. 7
Fig. 7 Validation of the JAK/STAT3-microtubule-RhoA axis in DLBCL in vivo and clinical samples. a A schema for showing the experimental design of the mice experiment. The HT-1 subline was injected into the spleen of SCID/beige mice. Before Taxol or Ruxolitinib injection, the primary tumors were removed. The drugs were injected intraperitoneally on Day 0 and bioluminescent signals were taken at the indicated time points. NaCl, vehicle control. n = 6 for each group. b Quantification of disseminated lesions (red dotted lines) in a. c Representative pictures of immunohistochemical staining of the Y705-phosphorylated STAT3, and S19-phosphorylated MCL, RhoH, and CD20 in DLBCL cases. The clinical stage of the representative cases is indicated. Scale bar = 25 μm. d Distribution of H scores of Y705-phosphorylated STAT3 in DLBCL patients with different stages. e Distribution of H scores of RhoH in DLBCL patients with different stages. *P < 0.05 by Student’s t-test. See also Supplementary Fig. 7CC BY · PubMed Central
Fig. 8 A schema for showing the molecular mechanism guiding cell movement in disseminated stage DLBCL
Fig. 8 A schema for showing the molecular mechanism guiding cell movement in disseminated stage DLBCLCC BY · PubMed Central

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Cited by

Metadata

Authors
Pan, Y.-R., Chen, C.-C., Chan, Y.-T., Wang, H.-J., Chien, F.-T., Chen, Y.-L., Liu, J.-L., & Yang, M.-H.
ORCID
Venue
Nature Communications
Publisher
Springer Science and Business Media LLC
Published
2018-09-12
Volume
9
Issue
1
ISSN
2041-1723
DOI
10.1038/s41467-018-06134-z
PMC
PMC6135800
Cited by
47
References
3
Language
en
Licence
Funders
  • Ministry of Science and Technology, Taiwan (104-2321-B-010-005, 104-0210-01-09-02, 107-0210-01-19-01)
  • National Health Research Institutes (NHRI-EX107-10622BI)
  • Ministry of Health and Welfare (MOHW107-TDU-B-211-114019)

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