1.Inhibition of hepatic stellate cell proliferation and activation by bone marrow mesenchymal stem cells in vitro
Chinese Journal of Pathophysiology 1989;0(06):-
AIM: To investigate the effect of bone marrow (BM)-derived mesenchymal stem cells (MSCs) on the proliferation of hepatic stellate cells (HSCs) in vitro. METHODS: MSCs and HSCs were isolated from BM and liver respectively in Fischer 344 rats. HSCs were re-cultured on plastic and appeared activated morphologically. The coculture system was set up by transwell insert with pore size 0.4 ?m and 6-well plates. The 5th passage HSCs were cultured on the 6-well plastic plate and MSCs or buffalo rat liver cells (BRLs) or HSCs seeded on the transwell insert. Smooth muscle ?-actin (?-SMA) and desmin expression of HSCs were tested by immunocytochemistry and quantified by IBAS 2.5 software. RESULTS: After coculture for 24 h, the proliferation of HSCs was inhibited to a moderate extent by MSCs. With the coculture time lasted, the significant growth suppression of HSCs was affected by MSCs (15.7% and 30.3%) but not BRL cells in 48 h and in 72 h. After coculture for 72 h, the expression of ?-SMA in HSCs was lower in MSCs coculture system than that with BRLs or HSCs alone system (50.2% vs 90.2%, 95.6%, P
2.Inhibition of invasion and metastasis by imrecoxib and its mechanisms in lung adenocarcinoma A549 cell xenograft in nude mice
Lingchan WANG ; Lijing CUI ; Dongchang WANG ; Yunxia ZHAO ; Zhifang ZHAO ; Gang CHEN
China Oncology 2017;27(1):1-6
Background and purpose:Cyclooxygenase-2 (COX-2) participates in angiogenesis and lymph node metastasis of lung cancer. COX-2 inhibitors could inhibit invasion and metastasis of lung cancer. This study aimed to investigate the inhibition of invasion and metastasis by COX-2 inhibitor imrecoxib in xenograft tumor of lung adenocar-cinoma A549 cell in nude mice and to explore its possible mechanisms, in addition, to observe the efficacy of imrecoxib combined with lobaplatin.Methods:Thirty male BALB/c nude mice were injected subcutaneously with A549 cells into the right axillary region to establish xenograft models. Twenty-nine successfully modeled mice were randomly divided into four groups: control group (n=7), imrecoxib group (n=8), lobaplatin group (n=7), imrecoxib combined with lobaplatin group (n=7). The control group was treated with the same amount of sterile distilled water and injected with the same amount of 0.9% sodium chloride solution via caudal vein. The treatment group was treated with imrecoxib tablets 40 mg/kg per day through gavage and injected with lobaplatin 7.5 mg/kg per week via caudal vein respectively. The diet, physical activity and other normal conditions of nude mice were observed everyday. After 6 weeks, 29 mice were sacrificed and transplanted tumor tissues were cut off. The expression of PTEN, cortactin protein and mRNA were detected by immunohistochemistry and real-time PCR. The data were analyzed with one-way anova and non-parametric test.Results:In the last week, the diet and physical activity of all nude mice were less than before, and they became thinner, which were more obvious among the mice in lobaplatin group and imrecoxib combined with lobaplatin group. Compared with the control group, the expression of PTEN protein and mRNA were significantly increased in imrecoxib group and imrecoxib combined with lobaplatin group (P<0.001, respectively). Compared with the control group, the expression of cortactin protein and mRNA were significantly decreased in imrecoxib group and imrecoxib combined with lobaplatin group (P<0.001, respectively). PTEN and cortactin protein, PTEN and cortactin mRNA had significantly negative correlation (r=-0.660, -0.983,P<0.001, respectively). Conclusion:Imrecoxib can inhibit non-small cell lung cancer invasion and metastasis which may be involved in upregu-lating PTEN protein and reducing cortactin protein. Imrecoxib could enhance the effect of lobaplatin chemotherapy.
3.Effects of cotransplantation of rat bone marrow mesenchymal stem cells and bone marrow on hematopoiesis
Junxia LEI ; Zhenyu GUO ; Dongchang ZHAO ; Hongxia LI ; Weihua YU ; Xiuming ZHANG ; Qin ZHENG ; Qing WEI ; Shunong LI ; Peng XIANG
Chinese Journal of Pathophysiology 1989;0(06):-
AIM: To investigate effects of rBMMSC on hematopoiesis and immune reconstitution after allo-hematopoietic stem cells transplantation (HSCT). METHODS: Allogeneic BMT model from Fischer344 rats (RT-1Al) to Wistar rats (RT-1Au) was established. The effects of MSCs on hematopoietic reconstitution and immune reconstitution were studied by observing the survival rate, peripheral blood counts, thymus counts, spleen counts, bone marrow counts and immune function analysis at 30 days after transplantation. RESULTS: 1. Cotransplantation of MSCs and bone marrow (BM) was demonstrated to improve hematopoietic reconstitution. Lymphocyte and platelet counts in peripheral blood in cotransplantation groups were higher than those in control groups. More bone marrow neucleated cells were also observed in cotransplantation groups. 2. Cotransplantation of MSCs and BM improved immune reconstitution. First, overall thymic cellularity and spleen cellularity significantly increased in cotransplantation groups at day 30. Secondly, cotransplantation improved immune functional recovery. Non-specific lymphocytes proliferation reaction induced by ConA and LPS increased in cotransplantation group, and so did for allogeneic mixed-lymphocyte reaction. CONCLUSION: Hematopoietic reconstitution and immune reconstitution were significantly enhanced by MSCs cotransplanted with BM.
4.Inhibitory effect of imrecoxib combined with lobaplatin on tumor growth and lymph node metastasis ;of human lung cancer xenografts in nude mice
Dongchang WANG ; Lingchan WANG ; Lingjiao WANG ; Gang CHEN ; Yunxia ZHAO ; Zhifang ZHAO ; Yahua LI
Chinese Journal of Oncology 2016;38(5):340-345
Objective To evaluate the inhibitory effect of imrecoxib combined with lobaplatin on tumor growth and lymph node metastasis of human lung adenocarcinoma xenografts in nude mice, and to explore its possible mechanisms. Methods Human lung cancer A549 cells were injected into Bal B/c nude mice subcutaneously. Twenty?eight healthy male nude mice were randomly divided into 4 groups:the control group, imrecoxib group, lobaplatin group and imrecoxib combined with lobaplatin group. Each group was treated with appropriate drugs and the tumor size was measured every five days. The expression of ezrin and E?cadherin protein was detected by immunohistochemistry and flow cytometry. Ezrin and E?cadherin mRNA were detected by real?time PCR. Results The tumor inhibition rates of imrecoxib group, lobaplatin group and combination group were 36.7%, 54.6% and 69.2%, respectively. The tumor volumes of imrecoxib group [(905.33±113.31) mm3] and combination group [(507.74±77.50) mm3] were significantly lower than that of the control group (1355.33±189.04) mm3(P<0.05), and the tumor weights were significantly reduced [(1.13±0.14) g, (0.63±0.10) g respectively] vs. (1.69±0.24) g (P<0.05). The expressions of ezrin protein and mRNA in the imrecoxib group and combined treatment group were significantly lower than that of the control group (136.53±35.52, 74.72±19.48 vs. 175.62±21.16 for protein expression level;0.54± 0.03, 0.36±0.03 vs. 1.02±0.02 for mRNA expression level, respectively, P<0.05 for both), while the expression of E?cadherin protein and mRNA in the imrecoxib group and combined treatment group was significantly higher than that of the control group ( 253. 78 ± 38. 87, 308. 94 ± 24. 67 vs. 213. 66 ± 30. 31 for protein expression level;2.19±0.02, 3.02±0.02 vs. 1.05±0.03 for mRNA expression level, respectively, P<0.05 for both) . There was a significant negative correlation between ezrin protein and E?cadherin protein (r=-0.737, P<0.01), as well as between ezrin mRNA and E?cadherin mRNA (r=-0.977, P<0.01). Conclusions Administration of imrecoxib combined with lobaphatin has inhibitory effects on the growth of non?small cell lung cancer xenografts and lymph node metastasis via down?regulated ezrin and upregulated E?cadherin. Imrecoxib and lobaplatin have a synergistic antitumor effect.
5.Inhibitory effect of imrecoxib combined with lobaplatin on tumor growth and lymph node metastasis ;of human lung cancer xenografts in nude mice
Dongchang WANG ; Lingchan WANG ; Lingjiao WANG ; Gang CHEN ; Yunxia ZHAO ; Zhifang ZHAO ; Yahua LI
Chinese Journal of Oncology 2016;38(5):340-345
Objective To evaluate the inhibitory effect of imrecoxib combined with lobaplatin on tumor growth and lymph node metastasis of human lung adenocarcinoma xenografts in nude mice, and to explore its possible mechanisms. Methods Human lung cancer A549 cells were injected into Bal B/c nude mice subcutaneously. Twenty?eight healthy male nude mice were randomly divided into 4 groups:the control group, imrecoxib group, lobaplatin group and imrecoxib combined with lobaplatin group. Each group was treated with appropriate drugs and the tumor size was measured every five days. The expression of ezrin and E?cadherin protein was detected by immunohistochemistry and flow cytometry. Ezrin and E?cadherin mRNA were detected by real?time PCR. Results The tumor inhibition rates of imrecoxib group, lobaplatin group and combination group were 36.7%, 54.6% and 69.2%, respectively. The tumor volumes of imrecoxib group [(905.33±113.31) mm3] and combination group [(507.74±77.50) mm3] were significantly lower than that of the control group (1355.33±189.04) mm3(P<0.05), and the tumor weights were significantly reduced [(1.13±0.14) g, (0.63±0.10) g respectively] vs. (1.69±0.24) g (P<0.05). The expressions of ezrin protein and mRNA in the imrecoxib group and combined treatment group were significantly lower than that of the control group (136.53±35.52, 74.72±19.48 vs. 175.62±21.16 for protein expression level;0.54± 0.03, 0.36±0.03 vs. 1.02±0.02 for mRNA expression level, respectively, P<0.05 for both), while the expression of E?cadherin protein and mRNA in the imrecoxib group and combined treatment group was significantly higher than that of the control group ( 253. 78 ± 38. 87, 308. 94 ± 24. 67 vs. 213. 66 ± 30. 31 for protein expression level;2.19±0.02, 3.02±0.02 vs. 1.05±0.03 for mRNA expression level, respectively, P<0.05 for both) . There was a significant negative correlation between ezrin protein and E?cadherin protein (r=-0.737, P<0.01), as well as between ezrin mRNA and E?cadherin mRNA (r=-0.977, P<0.01). Conclusions Administration of imrecoxib combined with lobaphatin has inhibitory effects on the growth of non?small cell lung cancer xenografts and lymph node metastasis via down?regulated ezrin and upregulated E?cadherin. Imrecoxib and lobaplatin have a synergistic antitumor effect.