ABSTRACT
AIM: To observe the effect of soluble glycoprotein 130(sgp130)on expression of p-STAT3 and vascular endothelial growth factor(VEGF)-A in retina of mice with diabetes mellitus(DM), and explore the possibility of sgp130 in interfering with inflammatory damage of diabetic retinopathy(DR).METHODS: A total of 45 mice were randomly divided into normal group, DM group and sgp130 group. DM models were made in DM group and sgp130 group with streptozotocin. No special intervention was given to normal group and DM group, but sgp130 group was given intravitreal injection of 1.5mg/mL sgp130 2μL at the 1 and 5wk. After 10wk, all the mice were sacrificed to assess the protein expression of interleukin 6(IL-6), p-STAT3 and VEGF-A in the retina.RESULTS: The expressions of IL-6, p-STAT3 and VEGF-A in retina of DM group were higher than those of normal group at 10wk(all P<0.01). The expression of p-STAT3 and VEGF-A in sgp130 group were lower than those in DM group(all P<0.01).CONCLUSION: The sgp130 can selectively antagonize the trans signal transduction pathway of IL-6, down-regulate the expression of downstream inflammatory factors VEGF-A, and it may be used in the intervention of retinal inflammatory damage related with IL-6 in DM.
ABSTRACT
@#目的：探究环状 RNA(circular RNA, circRNA）0072088 在非小细胞肺癌（non-small cell lung cancer，NSCLC）细胞中 的生物学功能及其作用机制。方法：在公共基因芯片数据库 Gene Expression Omnibus（GEO）中下载 GSE101684 数据集，通过 GEO2R 分析得到差异基因。通过 qPCR 检测 NSCLC 细胞 H165、H358、H460、H226 和 A549 细胞中 circ_0072088 的表达水平， 随后采用 CCK-8 法和 Transwell 小室法分别检测 circ_0072088 对 NSCLC 细胞增殖、迁移和侵袭的作用。通过 CircInteractome 和 TargetScan 数据库预测 circ_0072088 与 miR-545-3p、miR-545-3p 与 STAT3 之间的靶向关系，并通过双荧光素酶报告基因实 验和 RNA 结合蛋白免疫沉淀（RNA binding protein immunoprecipitation，RIP）实验验证 circ_0072088、miR-545-3p 与 STAT3 之 间的靶向关系。结果：circ_0072088 在 NSCLC 细胞系中的表达均显著上调（均 P<0.05）。过表达 circ_0072088 促进了 NSCLC 细胞的增殖、侵袭和迁移（均 P<0.05）；敲低 circ_0072088 抑制了 NSCLC 细胞的增殖、侵袭和迁移（均 P<0.05）。miR-545-3p 是 circ_0072088 的下游靶点，可以被 circ_0072088 吸附；STAT3 是 miR-545-3p 的靶基因，可以被 circ_0072088 间接正向调控。 结论：Circ_0072088 通过调节 miR-545-3p /STAT3 轴促进 NSCLC 细胞的增殖和转移。
ABSTRACT
Aim To establish gefitinib resistant epidermal growth factor receptor(EGFR) mutant lung cancer cell lines and to explore the changes of downstream signaling pathway of EGFR. Methods Gefitinib concentration gradient method was used to induce the establishment of H3255 and HCC827 resistant cell lines, and CCK8 assay was used to detect the proliferation of gefitinib resistant cell lines H3255/GR and HCC827/ GR. Western blot was used to detect the changes of EGFR downstream signals in H3255, HCC827, H3255/GR and HCC827/GR cells. Results The proliferation of H3255/GR and HCC827/GR cells was significantly lower than that of non-drug resistant cells. The phosphorylation signal molecules p-AKT, p-ERK1/2 and p-STAT3 of H3255/GR and HCC827/ GR drug-resistant cell lines were no significantly changed compared with their non-drug-resistant cell lines. There was no significant change in the expression of p-STAT3 and p-ERKl/2 in H3255/GR cells treated with gefitinib, but the expression of p-AKT was significantly down-regulated, while gefitinib only slightly inhibited the expression of p-ERKl/2 in drugresistant H3255/GR cells. In HCC827/GR cells, p-AKT and p-STAT3 were not inhibited by gefitinib, while p-ERKl/2 was moderately inhibited. Conclusion There are significant differences in the signal mechanism of drug resistance between H3255/GR and HCC827/GR cell lines.
ABSTRACT
Objective To explore whether the IL-6/STAT3 signaling pathway regulate the expression of high mobility group proteins1 (HMGB1) in intestinal mucosa of rats with sepsis through the cecum ligation puncture (CLP). Methods One hundred and twenty male SD rats were randomly(random number) divided into three groups: sham operation group (group S, n=40), CLP group(group C, n=40) and anti-IL-6 monoclonal antibody group (group T, n=40). Rats in group S only received the simple laparotomy;Rats in group C and group T were established as a rat model of sepsis using CLP; rats in group T received the intraperitoneal injection of anti-IL-6 monoclonal antibody at 1 h after CLP, while the same volume of sodium lactate ringer's solution was injected to rats in group S and group C. Ten rats in each group were sacrificed at 3, 12, 24 and 48 h, respectively, and intestinal mucosa specimens were collected for pathological examinations by HE staining. The protein expression of HMGB1 and IL-6 were detected by immunohistochemistry, STAT3-protein by Western blot.and the levels of diamine oxidase (DAO) and D lactic acid in plasma by spectrophotometric. Results Rats in group C and group T showed obvious intestinal damage to different degrees, significantly higher intestinal mucosa pathological scores and plasma levels of DAO and D-lactic acid compared with rats in group S (P<0.05). The protein expression of IL-6, HMGBl and p-STAT3 of intestinal mucosa in group C and group T also significantly increased compared with that in group S (P<0.05). The intestinal mucosa pathological score, plasma levels of DAO and D-lactic acid and protein expression of IL-6, HMGBl and STAT3 were decreased in group T compared with those in group C (P<0.05). The intestinal mucosa pathological scores were positively correlated with the protein expression of IL-6 and HMGB1 at 12, 24, and 48 h, respectively. Conclusions IL-6 and HMGBl were involved in the intestinal injury of septic rats. IL-6/STAT3 signaling pathway could up-regulate the expression of HMGB1 in intestinal mucosa of septic rats.
ABSTRACT
Objective: To observe the effect of herb-partition moxibustion on colonic inflammation in UC rats, and participate in the regulation of immune regulation via NF-κB pathway and STAT3 phosphorylation. Methods: Rats were randomly divided into normal group, the ulcerative colitis group (UC) group and herb-partition moxibustion group, with 8 rats in each group. The rat model of UC was induced by 4％ DSS. After successful modeling, the rats were treated with moxibustion on bilateral Tianshu acupoints (ST25) . Each acupoint was used with 2 Zhuang moxa, 1 time a day, for 7 times. The effect of the herb-partition moxibustion on UC rats was observed by comparing the histopathological and protein concentrations of serum proinflammatory cytokine. Western Blot was used to detect NF-κB pathway and STAT3 activity in colon tissue. Results: Colonic histopathology in the UC group showed that the mucosal epithelium with ulcer formation and obvious inflammatory response. The herb-partition moxibustion could repair colonic epithelial damage and reduce the inflammatory response of colon tissue in UC rats. Compared with the normal group, the serum IL-1β and IL-6 protein concentrations were significantly increased in the UC group (P ＜ 0.05, P ＜ 0.01), and the STAT3 phosphorylation level and protein expression levels of pIκB-α and NF-κB p65 were significantly increased. The protein expression level of IκB-α was significantly decreased. Compared with the UC group, the serum protein concentrations of IL-1β and IL-6 were significantly lower in the herb-partition moxibustion group (P ＜ 0.05), and protein expression level of NF-κB p65 was decreased in the colon tissue. The phosphorylation level of pIκB-α was decreased, while the protein expression level of IκB-α was increased. Conclusion: Herb-partition moxibustion reduced colonic inflammatory response in UC rats by DSS-induced, the underlying mechanism may related to decrease release of proinflammatory cytokines IL-1β and IL-6 via dual inhibition of NF-κB and STAT3 activation.
ABSTRACT
OBJECTIVE Although it is generally believed that nicotine accounts for the beneficial effect of smoking on ulcerative colitis, the underlying mechanisms remain not well understood. Our previous finding that nicotine inhibits inflammatory responses through inducing miRNA-124 prompted us to ask whether the miRNA is involved in the protective action of nicotine on UC. METHODS MiR-124 expres-sion in colon tissues and cells was determined by q-PCR and in situ hybridization.The effect of miR-124 on protective role of nicotine in ulcerative colitis was evaluated in DSS-treated mice and IL-6-treated Caco-2 colon epithelial cells. Expression of p-STAT3/STAT3 was detected by immunohistochemistry and Western-blot analysis. RESULTS miR-124 expression is upregulated in colon tissues from UC patients and DSS-induced colitis mice. Nicotine treatment further elevated miR-124 level in lympho-cytes isolated from human ulcerative colonic mucosa and ulcerative colon tissues from DSS mice,both in infiltrated lymphocytes and epithelial cells. Administration of nicotine also reduced weight loss, improved DAI and decreased HE score in DSS-induced colitis mice.Moreover,knockdown of miR-124 in vivo significantly diminished the beneficial effect of nicotine on murine colitis, and in vitro on IL-6-treated Caco-2 colon epithelial cells.Further analysis indicated that nicotine inhibited STAT3 activation in vivo and in IL-6-treated Caco-2 colon epithelial cells and Jurkat human T lymphocytes,in which miR-124 knockdown led to increased activation of STAT3. Blocking STAT3 activity alone is beneficial for DSS colitis and also abolished nicotine′s protective effect in this model.CONCLUSION These data indicated that nicotine exerts its protective action in UC through inducing miR-124 and its effect on STAT3, and suggest that the miR-124/STAT3 system is a potential target for the therapeutic intervention of UC.
ABSTRACT
OBJECTIVE Although it is generally believed that nicotine accounts for the beneficial effect of smoking on ulcerative colitis, the underlying mechanisms remain not well- understood. Our previous finding that nicotine inhibits inflammatory responses through inducing miRNA-124 prompted us to ask whether the miRNA is involved in the protective action of nicotine on UC. METHODS MiR-124 expression in colon tissues and cells was determined by q-PCR and in situ hybridization. The effect of miR-124 on protective role of nicotine in ulcerative colitis was evaluated in DSS-treated mice and IL-6-treated Caco-2 colon epithelial cells. Expression of p-STAT3/STAT3 was detected by immuno?histochemistry and Western blot analysis. RESULTS miR- 124 expression is upregulated in colon tissues from patients and DSS- induced colitis. Nicotine treatment further elevated miR- 124 level in colon tissues of the mice, in infiltrated lymphocytes and epithelial cells, and augmented miR- 124 expression in lymphocytes isolated from human ulcerative colon tissues. Administration of nicotine also reduced weight loss, improved DAI and decreased HE score in DSS-induced colitis. Moreover, knock?down of miR-124 in vivo significantly diminished the beneficial effect of nicotine, and in vitro on IL-6-treated Caco-2 colon epithelial cells. Further analysis indicated that nicotine inhibited STAT3 activation in vivo and in IL-6-treated Caco-2 colon epithelial cells and Jurkat human T lymphocytes, in which miR-124 knockdown led to increased activation of STAT3. CONCLUSION These data indicated that nicotine exerts its protective action in UC through inducing miR-124 and its effect on STAT3, suggesting that the miR-124/STAT3 system is a potential target for the therapeutic intervention of UC.
ABSTRACT
Objective To investigate expressions of STAT3 and phosphorylated STAT3(p-STAT3)in cutaneous malignant melanoma(CMM)lesions, and to explore their significance. Methods The expressions of STAT3 and p-STAT3 were detected with an immunohistochemical method using polymer enzyme-labeled secondary antibodies in skin specimens from 22 CMM lesions and 23 pigmented nevus lesions. The effects of STAT3 and p-STAT3 expressions on lymph node metastasis of CMM were evaluated. Results The expression rate of STAT3 was significantly higher in CMM lesions than in pigmented nevus lesions (95.5% vs. 56.5%, χ2 = 9.23, P 0.05), while the elevated expression of p-STAT3 in CMM lesions was a predictive factor for lymph node metastasis in CMM (OR = 1.88, 95% CI: 1.05 - 3.38, P < 0.05). Conclusion Increased expression of STAT3 and p-STAT3 in CMM may play important roles in the pathogenesis of CMM, and elevated p-STAT3 expression may contribute to invasion and metastasis of CMM.
ABSTRACT
Aim To investigate the inhibitory effect of Evodiamine on JAK2/STAT3 signal pathway in human colorectal cancer cell line HCT-116 . Methods Cells were cultured with 6. 0 μmol·L-1 Evodiamine for 2, 4 and 6 h, respectively. Cell nuclear morphology was detected by Hoechst staining and protein expression levels of JAK2 , p-JAK2 , STAT3 and p-STAT3 were examined by Western blot. Cells were treated with dif-ferent concentrations of AG490 for 48 h to select proper working concentration and cells treated with 6 μmol · L-1 EVO and 50 μmol · L-1 AG490 to compare the modulatory effect of EVO with AG490 on JAK2/STAT3 signal pathway. Results Hoechst staining revealed that Evodiamine could induce cells apoptosis, chroma-tin condensation gathered and typical apoptotic mor-phological changes in a time-dependent manner;West-ern Blot suggested that EVO could inhibit p-STAT3 significantly. After treatment with AG490, JAK2/STAT3 signal pathway was inactivated, the inhibitory effect of EVO on p-STAT3 was stronger than that of AG490 , while EVO combined with AG490 could fur-ther inhibit the expression of p-STAT3 significantly. Conclusions The anticancer effect of Evodiamine is mainly mediated by the modulation of JAK2/STAT3 signal pathway in HCT-116 cells.
ABSTRACT
The expressions of p-STAT3 and osteopontin in 22 cases of normal nevi and 43 cases of malignant melanoma were immunohistochemically detected,and the correlation between p-STAT3 and osteopontin in malignant melanoma and the correlations of p-STAT3 (or osteopontin) with invasion,metastasis and thickness of malignant melanoma were examined.The results showed p-STAT3 was expressed in 2 of 22 cases of normal nevi and 30 of 43 cases of malignant melanoma,while osteopontin was expressed in 3 cases of normal nevi and 29 cases of malignant melanoma.The expressions of p-STAT3 and osteopontin in melanoma were significantly higher than that in benign nevi.There existed significant correlations between the expression of p-STAT3 and that of osteopontin in melanoma.Furthermore,the expression rates of p-STAT3 were significantly higher in invasive or metastatic melanomas than that their non-invasive or non-metastatic counterparts,and the expression rates of osteopontin were significantly higher in invasive melanomas than that in non-invasive ones.It is concluded that p-STAT3 and osteopontin may play important roles in the pathogenesis of malignant melanoma.
ABSTRACT
Objective To investigate the effects of rapamycin on p-JAK2,STAT3,p-STAT3 and PCNA in ranal tissue of IgAN rats.Methods The animal models of IgA nephropathy were devided into control group,IgAN model group,losartan group and rapamycin group.The clinical efficacy of rapamycin,and its influences on the protein and mRNA expressions of STAT3,p-STAT3,p-JAK2 and PCNA were determined by western blot,RT-PCR and immunohistochemical techniques respectively.Results The protein and mRNA expressions of p-STAT3,STAT3,p-JAK2 and PCNA were significantly increased in kidney of IgAN rats(P