ABSTRACT
Objective:To investigate the difference in the radiation sensitivity of hematopoietic stem and progenitor cells (HSPCs) derived from fetal liver and bone marrow.Methods:HSPCs from fetal liver of 14.5 d embryo or bone marrow of 8 week-old mice were isolated to receive a single dose of 5 or 10 Gy irradiation in vitro using a 60Co irradiator. Twelve hours later, the cell apoptosis, mitochondrial reactive oxygen species (ROS) level, colony formation ability and DNA damage in HSPCs were detected. Freshly isolated HSPCs were injected into lethally irradiated CD45.1 + C57BL/6J mice (4.5 Gy+ 5 Gy with an interval of 30 min) Chimerism rate, lineage constitution, and cell cycle were analyzed 12 weeks after transplantation. Results:Compared with bone marrow HSPCs after irradiation, the percentage of apoptosis in fetal liver HSPCs was significantly higher ( t=16.21, 12.27, P<0.05), the level of ROS was dramatically elevated ( t=68.72, 18.89, P<0.05). At 10 Gy, fetal liver HSPCs could not form colonies at all ( t=12.41, 15.67, 9.46, P<0.05). γ-H2AX immunofluorescence staining showed that the DNA damage of fetal liver HSPCs was more severe after irradiation, and the number of Foci formed was significantly higher than that of bone marrow HSPCs ( t=2.27, 2.03, P< 0.05), which indicated that fetal liver HSPCs were more sensitive to radiation. The chimerism rate of transplanted fetal liver HSPCs was lower than that of bone marrow cells ( t=5.84, P<0.05) with a higher proportion of myeloid lineage, suggesting that fetal liver HSPCs had lower in vivo reconstitution capacity than bone marrow HSPCs and were more prone to myeloid differentiation. The cell cycle of bone marrow HSPCs from transplanted chimeric mice was examined, and the proportion of S-phase was significantly higher in the fetal liver group than that in the bone marrow group ( t=2.89, P<0.05). Mitochondrial stress results showed that fetal liver HSPCs had higher basal respiratory capacity ( t=39.19, P<0.05), proton leakage ( t=6.64, P<0.05), ATP production ( t=9.33, P<0.05), and coupling efficiency ( t=7.10, P<0.05) than bone marrow c-Kit + cells, while respiratory reserve capacity ( t=5.53, P< 0.05) was lower than that of bone marrow c-Kit + cells. Conclusions:HSPCs derived from fetal liver display higher radiosensitivty compared with bone marrow HSPCs, laying the foundation for an in-depth illustration of the effects of radiation on hematopoietic stem cells at different developmental stages.
ABSTRACT
Objective:To study the effect of different doses of 60Co γ-ray ionizing radiation on mitochondrial function in mouse hematopoietic stem and progenitor cells (HSPCs). Methods:C57BL/6 mice were divided into control group, 1 Gy irradiation group and 4.5 Gy irradiation group. The mitochondrial functions were detected at 12 h and 24 h after irradiation, including ROS level, membrane potential, mitochondrial structure, and mitochondrial stress. Bone marrow c-Kit + cells received a single 15 Gy irradiation in vitro, after 24 h, mitochondrial function was detected. Results:It was found that mice leukocytes ( t=12.41, 18.31, 16.48, 14.16, 19.08, 20.25, P<0.05), red blood cells ( t=4.81, 6.62, P<0.05) and platelets ( t=4.33, 6.68, P<0.05) were significantly reduced. The numbers of bone marrow colony formation unit ( t=16.27, 55.66, 17.06, 43.75, P<0.05), and HSPCs ( t=5.16, 11.55, P<0.05) were decreased dose-dependently post-irradiation. Under 1 Gy irradiation, the mitochondrial function and mitochondrial basal metabolic index of HSPCs ( t= 7.36, 3.68, 4.58, 3.15, 3.15, P<0.05) were enhanced at 24 h post-irradiation. Under 4.5 Gy irradiation, mitochondrial number, mitochondrial membrane potential ( t=12.29, 10.46, P<0.05), maximal respiration and spare respiratory capacity were decreased ( t=7.81, 5.78, 6.70, 5.83, P<0.05), ROS level was increased ( t=4.63, 4.12, P<0.05). The basal respiration and oxidative phosphorylated ATP production were reduced at 12 h after irradiation ( t=8.48, 3.80, P<0.05); and the proton leakage was increased ( t=6.57, P<0.05) and coupling efficiency was reduced ( t=11.43, P<0.05) at 24 h after irradiation. In cultured c-Kit + cells, the level of ROS ( t=11.30, P<0.05) and the maximum respiration and spare respiratory capacity were increased ( t=4.25, 3.44, P<0.05) while the mitochondrial membrane potential was decreased ( t=34.92, P<0.05) significantly. Conclusions:A method for systematically assessing mitochondrial function in HSPCs was established, and the effect of ionizing radiation on mitochondrial function of HSPCs was clarified, laying a foundation for further revealing the mechanism of ionizing radiation-induced mitochondrial damage in HSPCs.
ABSTRACT
Objective To compare the efficacy and safety of trastuzumab versus the combination of tras-tuzumab and lapatinib added to neoadjuvant chemotherapy in HER2-positive breast cancer. Methods We searched PubMed,MEDLINE,The Cochrane Library,Web of Science,CNKI,Wanfang datebase and the abstracts of major international conferences in recent 5 years to identify randomized controlled trials which met the inclusion and exclusion criteria. Study selection and analyses were undertaken according to the Cochrane Hand-book. Meta-analysis was performed using RevMan 5. 0 software. Results Four trials were identified with 779 eli-gible patients. The results of meta-analyses showed that the rate of pathological complete response was significant-ly higher in the group receiving rastuzumab and tlapatinib than that in the group with trastuzumab alone(53. 3%vs 38. 8% ,RR =1. 39,95 % CI:1. 20-1. 63;P < 0. 001). No statistical differences were observed in regarding adverse events among patients receiving trastuzumab or the combination of trastuzumab and lapatinib,except the grade Ⅲ-Ⅳ diarrhea(2. 2% vs 25. 6% ,RR =11. 54,95% CI:5. 69-23. 41;P <0. 001). Conclusion The com-bination of trastuzumab and lapatinib added to neoadjuvant chemotherapy in HER2-positive breast cancer is more effective,without more adverse reactions except diarrhea;it ia an effective and safe treatment.