核仁素与人骨肉瘤细胞化疗药物敏感性关系的实验研究
摘要
研究背景:骨肉瘤是一种好发于青少年的原发性高度恶性肿瘤,具有恶性程度高、肺转移早、致残率高、复发率高等临床特点。手术与新辅助化疗是目前治疗骨肉瘤的主要手段。虽然病人的5年生存率已得到显著提高,但多药耐药、肿瘤的复发与转移仍是骨肉瘤治疗的瓶颈问题。随着分子生物学的发展,基因治疗在骨肉瘤治疗中的作用逐渐被重视。
核仁素(又称C23)是目前发现的271种核仁蛋白质中含量最多的一种,主要定位于细胞核仁内,少量分布在胞浆和细胞膜表面。核仁素与肿瘤发生、发展相关的功能有:(1)促进细胞的增殖与生长;(2)抗凋亡作用,已被列为细胞凋亡相关蛋白中的一员;(3)通过与DNA或相关蛋白质的结合,在DNA复制、重组和修复中发挥调控作用,在不同的细胞环境中交替发挥原癌基因或抑癌基因的功能;(4)参与了肿瘤新生血管生成的过程。
目前国内外对核仁素的研究集中于基础和制药领域,还没有临床数据显示核仁素与骨肉瘤的关联,但核仁素广泛的生物学功能为其在骨肉瘤诊疗方面的应用研究提供了理论基础。本实验拟通过确认核仁素在骨肉瘤中的表达,采用反义寡核苷酸技术和转基因技术来探讨核仁素对顺铂诱导人骨肉瘤细胞株SaOS-2增殖、凋亡的影响,以阐明核仁素在基因治疗中的潜在意义,以期为寻找新的骨肉瘤早期诊断指标、预后评估指标及基因治疗方法提供线索。
第一部分C23在骨肉瘤组织中的表达
目的观察C23在骨肉瘤中的表达情况及其与年龄、性别、病理亚型和外科分期之间的关联。方法(1)对30例骨软骨瘤和30例骨肉瘤石蜡切片进行SABC法免疫组化染色,分析C23的阳性表达情况;用卡方检验分析C23的阳性表达与年龄、性别、病理亚型和外科分期之间的关联;(2)用Western Blot和RT-PCR方法检测C23蛋白、mRNA在新鲜骨肉瘤、骨软骨瘤组织中的表达;结果(1)在骨肉瘤组织中,C23核阳性表达率为90.0%,强表达50.0%;在骨软骨瘤组织中,C23核阳性表达率为53.3%,无强表达,两者差异显著(P＜0.01)。核仁素在骨肉瘤细胞核中的阳性表达与Enneking分期有关(P＜0.05),与其他三个因素无明显关系。(2)C23蛋白和mRNA在骨肉瘤和骨软骨瘤组织中的表达量有显著差异(P＜0.01)。结论核仁素在大多数骨肉瘤组织中高表达;核仁素在骨肉瘤组织中阳性表达率与肿瘤的恶性程度呈正相关。
第二部分C23表达下调对CDP诱导人骨肉瘤细胞株SaOS-2增殖抑制和凋亡的影响
目的观察C23表达下调对CDP诱导人骨肉瘤细胞株SaOS-2增殖抑制和凋亡的影响方法(1)用MTT试验检测并作图得CDP作用SaOS-224小时的IC50值,取合适值作为实验浓度;(2)设计合成C23正义、反义与随机寡核苷酸并转染SaOS-2细胞,用western Blot和RT-PCR检测C23反义寡核苷酸对C23蛋白和mRNA表达的影响;(3)将细胞随机分为五组:对照组、单纯用CDP组、转染正义寡核苷酸后再用CDP组、转染反义寡核苷酸后再用CDP组与转染随机寡核苷酸后再用CDP组;(4)用MTT法检测各组SaOS-2细胞的增殖率,并用Western Blot和RT-PCR检测各组C23、Bcl-2和Bax蛋白和mRNA的表达情况;(5)用DAPI染色方法和流式细胞术检测各组细胞凋亡核百分率、细胞凋亡率。结果(1)CDP24小时的IC50为12.32ug/ml,取10ug/ml作为实验浓度;(2)C23反义寡核苷酸显著抑制了C23蛋白和mRNA的表达,可使其表达水平下调,与对照组有显著差异(P＜0.01);(3)转染反义寡核苷酸后再用CDP组与单纯用CDP组比较,细胞增殖率下降了约18%(P＜0.01),C23与Bcl-2蛋白和mRNA表达均降低,但Bax表达则增高(P＜0.01);(4)转染反义寡核苷酸后再用CDP组与单纯用药组比较,细胞凋亡核百分率与细胞凋亡率均增高,有统计学差异(P＜0.01)。结论C23表达下调能促进CDP诱导的SaOS-2细胞增殖抑制和凋亡。
第三部分C23过表达对ODP诱导人骨肉瘤细胞株SaOS-2增殖抑制和凋亡的影响
目的观察C23过表达对CDP诱导人骨肉瘤细胞株SaOS-2增殖抑制和凋亡的影响方法(1)构建pRSET-C23重组质粒并行测序鉴定;(2)将pRSET-C23重组质粒导入SaOS-2细胞,用Western Blot和RT-PCR方法检测其对C23蛋白和mRNA表达的影响;(3)将细胞随机分为四组:对照组、单纯用CDP组、转染空载体后再用CDP组与转染C23重组质粒后再用CDP组;(4)用MTT法检测各组SaOS-2细胞增殖率,并用Western Blot和RT-PCR方法检测各组C23、Bcl-2与Bax蛋白和mRNA的表达情况;(5)用DAPI染色方法和流式细胞术检测各组细胞凋亡核百分率、细胞凋亡率。结果(1)pRSET-C23重组质粒构建成功;(2)与转染空载体组比较,转染质粒组C23蛋白和mRNA表达明显升高,有统计学差异(P＜0.01);(3)转染C23重组质粒后再用CDP组的SaOS-2细胞,其增殖率与单纯用CDP组比较,升高了约16%(P＜0.01),C23与Bcl-2的蛋白和mRNA表达均增高,但Bax表达则降低(P＜0.01);(4)转染C23重组质粒后再用CDP组的SaOS-2细胞,与单纯用CDP组比较,细胞凋亡核百分率、细胞凋亡率均降低,有统计学差异(P＜0.01)。结论C23过表达能抑制CDP诱导的SaOS-2细胞增殖抑制和凋亡。
Background:Osteosarcoma is the most common high-grade primary bone tumor with a high degree of early lung metastasis and a high rate of deformity.It occurs in adolescents and children.Surgery and neoadjuvant chemotherapy are main treatments of it.Though the two ways has improved cure rates remarkably,the frequent acquisition of drug-resistant phenotypes and the occurrence of second malignancies are major barriers to the successful treatment of osteosarcoma.With the development of molecular Biology,gene therapy was used more often to heal it.
C23 is one of the most abundant nonribosomal proteins of the nucleolus It is found almost exclusively within the nucleolus,but increasing evidence indicates its presence at the surface and in cytoplasm of various cell types.The following functions of C23 have relationship with the occurrence and development of tumor:(1) C23 is a major actor in promoting cell proliferation.Its amount is correlated with the proliferative status of the cell:C23 level is higher in tumours so that it has been described as a diagnostic marker.(2) C23 has been identified as one of apoptosis-related proteins according to its anti-apoptotic role;(3) C23 can support roles in DNA replication,repair and recombination.It could function either as a "caretaker",a "gatekeeper" or a "proto-oncogene", depending on the cellular context.(4) C23 participate the process of neovessels formn.
In Persent study,to clarify the potential sense of C23 in Gene-therapy of osteosarcoma and to provide an experimental clue and academic evidence,firstly we will affirm the expression of C23 in osteosarcoma. Seeondly,using antisense technology and transgenic technology we are going to probe into the relationship of C23 with osteosarcoma and to observe the effect of C23 on inhibition of proliferation and apoptosis induced by cisplatin in human osteosarcoma SaOS-2 cells.The effect of C23 may shed new light on the cancer therapy and become a beneficial supplement for osteosarcomar traditional therapy.
PartⅠStudy on the expression of C23 in osteosarcoma tissue
Objective To investigate the expression of C23 in osteosarcoma and its correlation with clinical features.Methods(1)The expression of C23 was evaluated by immunohistochemistry in paraffin-embedded tissues from 30 cases of osteosarcoma and 30 cases of osteochondroma;The correlation between the expression of C23 and its clinical features was analyzed accordingly.(2) The expression of C23 protein and mRNA were detected by Immunoblotting and RT-PCR in fresh tissues of osteosarcoma and osteochondroma.Results(1) The C23 expression showed significant difference between osteosarcoma and osteochondroma(P＜0.01).The positive rate of C23 in osteosacroma was 90.0%compared with 53.3%in osteochondroma.The positive expression of C23 was correlated with Enneking surgical staging.Its positive expression rate in Enneking surgical stagingⅠ-ⅡA was significantly lower than that in stagingⅡB-Ⅲ(P＜0.05). (2) The expression of C23 protein and mRNA were stronger in Osteosarcoma than in osteochondroma(P＜0.01)。Conclusion A high expression of C23 was found in osteosarcoma tissue.And the expression of nucleolin has positive correlation with malignant degree.
PartⅡstudy on the effect of C23 down-regulation on inhibition of proliferation and apoptosis induced by cisplatin in human osteosarcoma SaOS-2 cells
Objective To investigate the effect of C23 down-regulation on inhibition of proliferation and apoptosis induced by cisplatin in human osteosarcoma SaOS-2 cells.Methods(1) SaOS-2 cells were treated with different concentrations of CDP,then cell viability was analyzed with MTT assay;Growth curve was drawn to get a suitable concentration as experimental concentration;(2) Targeted SODN、RODN and ASODN of C23 were designed and synthesized,then transfected to osteosarcoma SaOS-2 cells;The expression of C23 protein and mRNA were detected by Immunoblotting and RT-PCR.(3) SaOS-2 cells were divided to five groups randomly:control group;CDP group;SODN+CDP group; ASODN+ CDP group and RODN+CDP group;(4) Cell growth suppression of each group was analyzed by MTT assay.RT-PCR and Immunoblotting were used to detected the expression of C23、Bcl-2 and Bax mRNA and protein in each group;(5) Morphologic character of apoptosis was evaluated by DAPI staining;The percentage of apoptotic cell was analyzed by flowcytometer assay.Results(1) The IC50 of CDP was 12.32ug/ml and 10ug/ml was used as experimental concentration;(2) ASODN can inhibit remarkably the expression of C23 mRNA and protein in SaOS-2 cells(P＜0.01);(3) The proliferation of SaOS-2 cells was inhibited remarkably in ASODN+CDP group compared with CDP group (P＜0.01).The expression of C23、Bcl-2 mRNA and protein were weaker and that of Bax mRNA and protein were stronger in ASODN+CDP group compared with CDP group(P＜0.01).(4) C23 antisense oligonucleotides could promote cisplatin-mediated apoptosis in SaOS-2 cells with a higher percentgae of apoptotic cells than CDP group.(P＜0.01).Conclusion Those results suggested that down-regulation of C23 could contribute to cisplatin-mediated inhibition of proliferation and apoptosis in SaOS-2 cells.
PartⅢStudy on the effect of C23 over-expression on inhibition of proliferation and apoptosis induced by cisplatin in human osteosarcoma SaOS-2 cells
Objective To investigate the effect of C23 over-expression on inhibition of proliferation and apoptosis induced by cisplatin in human osteosarcoma SaOS-2 cells.Methods(1) C23 recombinant plasmid was constructed using gene engineering and was tested by sequence analysis; (2) The expression of C23 protein and mRNA in SaOS-2 cells transfected by C23 recombinant plasmid were detected by Immunoblotting and RT-PCR;(3) SaOS-2 cells were divided to four groups randomly:Control group;CDP group;V(vector) +CDP group and P(plasmid)+CDP group;(4) Cell growth suppression of each group was analyzed by MTT assay.RT-PCR and Immunoblotting were used to detected the expression of C23、Bcl-2 and Bax mRNA and protein in each group;(5) Morphologic character of apoptosis was evaluated by DAPI staining;The percentage of apoptotic cell was analyzed by flowcytometer assay.Results(1)C23 recombinant plasmid was constructed successfully;(2) Expression of C23 mRNA and protein was higher in SaOS-2 cells transfected with C23 recombinant plasmid than SaOS-2 cells transfected with pRSET-B vector.(P＜0.01);(3) C23 over-expression could block significantly the inhibition of proliferation induced by cisplatin, there had statistic difference between P+CDP group and CDP gorup(P＜0.01).The expression of C23 and Bcl-2 mRNA and protein were stronger and that of Bax mRNA and protein were weaker in P+CDP group than in CDP group(P＜0.01);(4) Over expression of C23 could significantly inhibit cisplatin-mediated apoptosis in SaOS-2 cells with a lower percentage of apoptotic cells compared with CDP group(P＜0.05) Conclusion Over-expression of C23 could increase resistance of SaOS-2 cells to cisplatin.
核仁素(又称C23)是目前发现的271种核仁蛋白质中含量最多的一种,主要定位于细胞核仁内,少量分布在胞浆和细胞膜表面。核仁素与肿瘤发生、发展相关的功能有:(1)促进细胞的增殖与生长;(2)抗凋亡作用,已被列为细胞凋亡相关蛋白中的一员;(3)通过与DNA或相关蛋白质的结合,在DNA复制、重组和修复中发挥调控作用,在不同的细胞环境中交替发挥原癌基因或抑癌基因的功能;(4)参与了肿瘤新生血管生成的过程。
目前国内外对核仁素的研究集中于基础和制药领域,还没有临床数据显示核仁素与骨肉瘤的关联,但核仁素广泛的生物学功能为其在骨肉瘤诊疗方面的应用研究提供了理论基础。本实验拟通过确认核仁素在骨肉瘤中的表达,采用反义寡核苷酸技术和转基因技术来探讨核仁素对顺铂诱导人骨肉瘤细胞株SaOS-2增殖、凋亡的影响,以阐明核仁素在基因治疗中的潜在意义,以期为寻找新的骨肉瘤早期诊断指标、预后评估指标及基因治疗方法提供线索。
第一部分C23在骨肉瘤组织中的表达
目的观察C23在骨肉瘤中的表达情况及其与年龄、性别、病理亚型和外科分期之间的关联。方法(1)对30例骨软骨瘤和30例骨肉瘤石蜡切片进行SABC法免疫组化染色,分析C23的阳性表达情况;用卡方检验分析C23的阳性表达与年龄、性别、病理亚型和外科分期之间的关联;(2)用Western Blot和RT-PCR方法检测C23蛋白、mRNA在新鲜骨肉瘤、骨软骨瘤组织中的表达;结果(1)在骨肉瘤组织中,C23核阳性表达率为90.0%,强表达50.0%;在骨软骨瘤组织中,C23核阳性表达率为53.3%,无强表达,两者差异显著(P＜0.01)。核仁素在骨肉瘤细胞核中的阳性表达与Enneking分期有关(P＜0.05),与其他三个因素无明显关系。(2)C23蛋白和mRNA在骨肉瘤和骨软骨瘤组织中的表达量有显著差异(P＜0.01)。结论核仁素在大多数骨肉瘤组织中高表达;核仁素在骨肉瘤组织中阳性表达率与肿瘤的恶性程度呈正相关。
第二部分C23表达下调对CDP诱导人骨肉瘤细胞株SaOS-2增殖抑制和凋亡的影响
目的观察C23表达下调对CDP诱导人骨肉瘤细胞株SaOS-2增殖抑制和凋亡的影响方法(1)用MTT试验检测并作图得CDP作用SaOS-224小时的IC50值,取合适值作为实验浓度;(2)设计合成C23正义、反义与随机寡核苷酸并转染SaOS-2细胞,用western Blot和RT-PCR检测C23反义寡核苷酸对C23蛋白和mRNA表达的影响;(3)将细胞随机分为五组:对照组、单纯用CDP组、转染正义寡核苷酸后再用CDP组、转染反义寡核苷酸后再用CDP组与转染随机寡核苷酸后再用CDP组;(4)用MTT法检测各组SaOS-2细胞的增殖率,并用Western Blot和RT-PCR检测各组C23、Bcl-2和Bax蛋白和mRNA的表达情况;(5)用DAPI染色方法和流式细胞术检测各组细胞凋亡核百分率、细胞凋亡率。结果(1)CDP24小时的IC50为12.32ug/ml,取10ug/ml作为实验浓度;(2)C23反义寡核苷酸显著抑制了C23蛋白和mRNA的表达,可使其表达水平下调,与对照组有显著差异(P＜0.01);(3)转染反义寡核苷酸后再用CDP组与单纯用CDP组比较,细胞增殖率下降了约18%(P＜0.01),C23与Bcl-2蛋白和mRNA表达均降低,但Bax表达则增高(P＜0.01);(4)转染反义寡核苷酸后再用CDP组与单纯用药组比较,细胞凋亡核百分率与细胞凋亡率均增高,有统计学差异(P＜0.01)。结论C23表达下调能促进CDP诱导的SaOS-2细胞增殖抑制和凋亡。
第三部分C23过表达对ODP诱导人骨肉瘤细胞株SaOS-2增殖抑制和凋亡的影响
目的观察C23过表达对CDP诱导人骨肉瘤细胞株SaOS-2增殖抑制和凋亡的影响方法(1)构建pRSET-C23重组质粒并行测序鉴定;(2)将pRSET-C23重组质粒导入SaOS-2细胞,用Western Blot和RT-PCR方法检测其对C23蛋白和mRNA表达的影响;(3)将细胞随机分为四组:对照组、单纯用CDP组、转染空载体后再用CDP组与转染C23重组质粒后再用CDP组;(4)用MTT法检测各组SaOS-2细胞增殖率,并用Western Blot和RT-PCR方法检测各组C23、Bcl-2与Bax蛋白和mRNA的表达情况;(5)用DAPI染色方法和流式细胞术检测各组细胞凋亡核百分率、细胞凋亡率。结果(1)pRSET-C23重组质粒构建成功;(2)与转染空载体组比较,转染质粒组C23蛋白和mRNA表达明显升高,有统计学差异(P＜0.01);(3)转染C23重组质粒后再用CDP组的SaOS-2细胞,其增殖率与单纯用CDP组比较,升高了约16%(P＜0.01),C23与Bcl-2的蛋白和mRNA表达均增高,但Bax表达则降低(P＜0.01);(4)转染C23重组质粒后再用CDP组的SaOS-2细胞,与单纯用CDP组比较,细胞凋亡核百分率、细胞凋亡率均降低,有统计学差异(P＜0.01)。结论C23过表达能抑制CDP诱导的SaOS-2细胞增殖抑制和凋亡。
Background:Osteosarcoma is the most common high-grade primary bone tumor with a high degree of early lung metastasis and a high rate of deformity.It occurs in adolescents and children.Surgery and neoadjuvant chemotherapy are main treatments of it.Though the two ways has improved cure rates remarkably,the frequent acquisition of drug-resistant phenotypes and the occurrence of second malignancies are major barriers to the successful treatment of osteosarcoma.With the development of molecular Biology,gene therapy was used more often to heal it.
C23 is one of the most abundant nonribosomal proteins of the nucleolus It is found almost exclusively within the nucleolus,but increasing evidence indicates its presence at the surface and in cytoplasm of various cell types.The following functions of C23 have relationship with the occurrence and development of tumor:(1) C23 is a major actor in promoting cell proliferation.Its amount is correlated with the proliferative status of the cell:C23 level is higher in tumours so that it has been described as a diagnostic marker.(2) C23 has been identified as one of apoptosis-related proteins according to its anti-apoptotic role;(3) C23 can support roles in DNA replication,repair and recombination.It could function either as a "caretaker",a "gatekeeper" or a "proto-oncogene", depending on the cellular context.(4) C23 participate the process of neovessels formn.
In Persent study,to clarify the potential sense of C23 in Gene-therapy of osteosarcoma and to provide an experimental clue and academic evidence,firstly we will affirm the expression of C23 in osteosarcoma. Seeondly,using antisense technology and transgenic technology we are going to probe into the relationship of C23 with osteosarcoma and to observe the effect of C23 on inhibition of proliferation and apoptosis induced by cisplatin in human osteosarcoma SaOS-2 cells.The effect of C23 may shed new light on the cancer therapy and become a beneficial supplement for osteosarcomar traditional therapy.
PartⅠStudy on the expression of C23 in osteosarcoma tissue
Objective To investigate the expression of C23 in osteosarcoma and its correlation with clinical features.Methods(1)The expression of C23 was evaluated by immunohistochemistry in paraffin-embedded tissues from 30 cases of osteosarcoma and 30 cases of osteochondroma;The correlation between the expression of C23 and its clinical features was analyzed accordingly.(2) The expression of C23 protein and mRNA were detected by Immunoblotting and RT-PCR in fresh tissues of osteosarcoma and osteochondroma.Results(1) The C23 expression showed significant difference between osteosarcoma and osteochondroma(P＜0.01).The positive rate of C23 in osteosacroma was 90.0%compared with 53.3%in osteochondroma.The positive expression of C23 was correlated with Enneking surgical staging.Its positive expression rate in Enneking surgical stagingⅠ-ⅡA was significantly lower than that in stagingⅡB-Ⅲ(P＜0.05). (2) The expression of C23 protein and mRNA were stronger in Osteosarcoma than in osteochondroma(P＜0.01)。Conclusion A high expression of C23 was found in osteosarcoma tissue.And the expression of nucleolin has positive correlation with malignant degree.
PartⅡstudy on the effect of C23 down-regulation on inhibition of proliferation and apoptosis induced by cisplatin in human osteosarcoma SaOS-2 cells
Objective To investigate the effect of C23 down-regulation on inhibition of proliferation and apoptosis induced by cisplatin in human osteosarcoma SaOS-2 cells.Methods(1) SaOS-2 cells were treated with different concentrations of CDP,then cell viability was analyzed with MTT assay;Growth curve was drawn to get a suitable concentration as experimental concentration;(2) Targeted SODN、RODN and ASODN of C23 were designed and synthesized,then transfected to osteosarcoma SaOS-2 cells;The expression of C23 protein and mRNA were detected by Immunoblotting and RT-PCR.(3) SaOS-2 cells were divided to five groups randomly:control group;CDP group;SODN+CDP group; ASODN+ CDP group and RODN+CDP group;(4) Cell growth suppression of each group was analyzed by MTT assay.RT-PCR and Immunoblotting were used to detected the expression of C23、Bcl-2 and Bax mRNA and protein in each group;(5) Morphologic character of apoptosis was evaluated by DAPI staining;The percentage of apoptotic cell was analyzed by flowcytometer assay.Results(1) The IC50 of CDP was 12.32ug/ml and 10ug/ml was used as experimental concentration;(2) ASODN can inhibit remarkably the expression of C23 mRNA and protein in SaOS-2 cells(P＜0.01);(3) The proliferation of SaOS-2 cells was inhibited remarkably in ASODN+CDP group compared with CDP group (P＜0.01).The expression of C23、Bcl-2 mRNA and protein were weaker and that of Bax mRNA and protein were stronger in ASODN+CDP group compared with CDP group(P＜0.01).(4) C23 antisense oligonucleotides could promote cisplatin-mediated apoptosis in SaOS-2 cells with a higher percentgae of apoptotic cells than CDP group.(P＜0.01).Conclusion Those results suggested that down-regulation of C23 could contribute to cisplatin-mediated inhibition of proliferation and apoptosis in SaOS-2 cells.
PartⅢStudy on the effect of C23 over-expression on inhibition of proliferation and apoptosis induced by cisplatin in human osteosarcoma SaOS-2 cells
Objective To investigate the effect of C23 over-expression on inhibition of proliferation and apoptosis induced by cisplatin in human osteosarcoma SaOS-2 cells.Methods(1) C23 recombinant plasmid was constructed using gene engineering and was tested by sequence analysis; (2) The expression of C23 protein and mRNA in SaOS-2 cells transfected by C23 recombinant plasmid were detected by Immunoblotting and RT-PCR;(3) SaOS-2 cells were divided to four groups randomly:Control group;CDP group;V(vector) +CDP group and P(plasmid)+CDP group;(4) Cell growth suppression of each group was analyzed by MTT assay.RT-PCR and Immunoblotting were used to detected the expression of C23、Bcl-2 and Bax mRNA and protein in each group;(5) Morphologic character of apoptosis was evaluated by DAPI staining;The percentage of apoptotic cell was analyzed by flowcytometer assay.Results(1)C23 recombinant plasmid was constructed successfully;(2) Expression of C23 mRNA and protein was higher in SaOS-2 cells transfected with C23 recombinant plasmid than SaOS-2 cells transfected with pRSET-B vector.(P＜0.01);(3) C23 over-expression could block significantly the inhibition of proliferation induced by cisplatin, there had statistic difference between P+CDP group and CDP gorup(P＜0.01).The expression of C23 and Bcl-2 mRNA and protein were stronger and that of Bax mRNA and protein were weaker in P+CDP group than in CDP group(P＜0.01);(4) Over expression of C23 could significantly inhibit cisplatin-mediated apoptosis in SaOS-2 cells with a lower percentage of apoptotic cells compared with CDP group(P＜0.05) Conclusion Over-expression of C23 could increase resistance of SaOS-2 cells to cisplatin.
引文
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