摘要
目的 观察不同浓度重组甲状旁腺激素1～34（简称hPTH）对成骨肉瘤细胞系SaOS-2细胞（简称SaOS-2细胞）骨碱性磷酸酶（ALP）和骨钙素（BGP）mRNA表达的影响.方法 SaOS-2细胞正常传代培养,用0、1、10、100 nmol/L hPTH处理细胞,分别在12、24、48 h提取细胞总RNA,反转录合成cDNA,采用实时荧光定量PCR的方法测定ALP和BGP mRNA的表达.结果 ①hPTH的不同剂量、不同的作用时间以及二者的交互作用对ALP的表达量均有影响（F值分别为29.32、2.92、7.64,P均〈0.05）.干预48 h,0、1、10、100 nmol/L组ALP mRNA的表达量（0.78±0.43、0.71±0.05、0.75±0.19、0.76±0.14）均低于同浓度组的12、24 h（12 h：1.01±0.16、1.37±0.38、1149±0.16、2.52±0.70,24 h：1.80±0.47、1.30±0.36、1.27±0.17、1.17±0.11,P均〈0.05）.干预12 h,100 nmol/L组ALP mRNA的表达量高于0 nmol/L组（P〈0.05）;干预24 h,1、10、100nmol/L组SaOS-2细胞ALP mRNA的表达量均低于0 nmol/L组（P均〈0.05）.②hPTH的不同剂量、不同的作用时间以及二者的交互作用对SaOS-2细胞BGP mRNA的表达量均有影响（F值分别为8.26、10.33、5.51,P均〈0.05）.干预48 h,0、1、10、100 nmol/L组BGP mRNA的表达量（1.17±0.28、0.98±0.08、0.92±0.17、0.84±0.59）均低于同浓度组的12、24 h（12 h：1.01±0.14、1.21±0.18、1.34±0.30、1.68±0.62,24 h：1.71±0.35、1141±0.47、1.28±0.31、1.01±0.18,P均〈0.05）.干预12 h,100 nmol/L组BGP mRNA的表达量高于0、1nmol/L组（P均〈0.05）;干预24 h,10、100 nmol/L组SaOS-2细胞BGP mRNA的表达量均低于0 nmol/L组（P均〈0.05）,100 nmol/L组SaOS-2细胞BGP mRNA的表达量低于1 nmoL/L组（P〈0.05）;干预48 h,10、100 nmol/L组SaOS-2细胞BGP mRNA的表达量均低于0 nmol/L组（P均〈0.05）.结论 在体外培养条件下,hPTH在短时间作用下可以显著增强SaOS-2细胞的成骨活性,随着刺激时间的延长,可使成骨活性呈现下降的趋势.
Objective To observe the effects of recombinant human parathyroid hormone 1 to 34（referred to as hPTH） on the expression level of alkaline phosphatase（ALP） and bone gla protein（BCP） in human osteosarcoma cell line SaOS-2（referred to as SaOS-2 cells）. Methods SaOS-2 cells were subcultured and treated with 1, 10 and 100 nmol/L hPTH for 12, 24 and 48 h. Total cellular RNA was extracted, cDNA was synthesized by reverse doses of hPTH, different duration of action, and their interaction on the expression level of ALP mRNA of SaOS-2 cells was significantly different（F = 29.32, 2.92, 7.64, all P 〈 0.05）. The expression level of ALP mRNA（0.78 ± 0.43, 0.71 ± 0.05, 0.75 ± 0.19, 0.76 ± 0.14） of SaOS-2 cells after treatment with 0, 1, 10 and 100 nmol/L hPTH for 48 h was lower than those of treated for 12 h（1.01 ± 0.16, 1.37 ± 0.38, 1.49 ± 0.16, 2.52 ± 0.70, all P〈 0.05） and 24 h （1.80 ± 0.47, 1.30 ± 0.36, 1.27 ± 0.17, 1.17 ± 0.11, all P〈 0.05）. The expression level of ALP mRNA of SaOS-2 cells after treatment with 100 nmol/L hPTH for 12 hours was higher than that of the control（P 〈 0.05）; the expression level of ALP mRNA of SaOS-2 cells after treatment with 1, 10 and 100 nmol/L hPTH for 24 h interaction on the expression level of BGP mRNA of SaOS-2 were significantly different （F = 8.26, 10.33, 5.51, all P〈 0.05）. The expression level of BGP mRNA（1.17 ± 0.28, 0.98 ± 0.08, 0.92 ± 0.17 and 0.84 ± 0.59） of SaOS2 cells after treatment with 0, 1, 10 and 100 nmol/L hPTH for 48 h was lower than those of treated for 12 h（ 1.01 ± 0.14, 1.21 ± 0.18, 1.34 ± 0.30, 1.68 ± 0.62, all P〈 0.05）, and 24 h（1.71 ± 0.35, 1.41 ± 0.47, 1.28 ± 0.31 and 1.01 ± 0.18, all P 〈 0.05）. The expression level of BGP mRNA of SaOS-2 cells after treatment with 100 nmol/L hPTH for 12 h was higher than that of those groups treated with 0 and 1 nmol/L hPTH（all P〈 0.05）. The expression level of BGP mRNA of SaOS-2 cells after treatment with 10 and 100 nmol/L hPTH for 24 h and 48 h was lower than those of the control（all P 〈 0.05）. The expression level of BGP mRNA of SaOS-2 cells after treatment with 100 nmol/L hPTH for 24 hours was lower than that the group treated with 1 nmol/L hPTH（P 〈 0.05）. Conclusions In vitro, hPTH significantly enhances osteogenic activities of human osteoblast in a short time, however, with prolonged stimulation time, osteogenic activity can show a downward trend.
出处
《中国地方病学杂志》
CAS
CSCD
北大核心
2011年第3期284-288,共5页
Chinese Jouranl of Endemiology
基金
基金项目：国家自然科学基金（30800956）
黑龙江省教育厅课题（11531098）
黑龙江省卫生厅课题（2006-288）
