CAJ | 학술논문

目的研究阿霉素干预胃癌细胞后PTEN的表达及其在阿霉素诱导胃癌细胞凋亡中的意义.方法 (1)阿霉索干预胃癌细胞BGC-823后以四甲基偶氮唑盐比色法(MTT法)和流式细胞法检测细胞存活率及凋亡率,并检测VFEN的mRNA和蛋白水平.(2)构建胃癌裸鼠异位种植瘤,采用原位末端标记(TUNEL)法检测异位种植瘤中胃癌细胞的凋亡情况,并用RT-PCR和Western blot法检测PTEN mRNA和蛋白的表达水平.(3)以PTEN特异性小干扰RNA(siRNA)转染BGC-823细胞,并以阿霉素进行干预,检测BGC-823细胞的存活率和凋亡率以及PTEN蛋白表达水平.结果 (1)阿霉素干预后,BGC-823细胞的生存率呈时间依赖性降低.(2)阿霉素能够有效诱导BGC-823细胞凋亡.(3)阿霉素在BGC-823细胞中可时间依赖性地促进PTEN的mRNA和蛋白水平的升高.裸鼠异位种植瘤试验中,阿霉素干预组的凋亡率[(28.11±1.05)%]明显高于对照组[(2.78±1.63)%];阿霉素干预组瘤体组织中PTEN mRNA和蛋白水平亦高于对照组(0.5667±0.0043比0.2217±0.0063,0.14±0.26比0.04±0.15,P值均<0.05).(4)转染与未转染[WEN siRNA的胃癌细胞以阿霉素干预后,PTEN siRNA转染组的PTEN蛋白表达水平明显低于对照组(P<0.0001),且PTEN siRNA转染组[(10.35±1.04)%]凋亡率明显小于未转染组[(31.37±3.58)%],P<0.05.结论阿霉素干预胃癌细胞后可以抑制其生长,诱导细胞凋亡,PTEN表达水平的升高可能是其机制之一.
목적 연구아매소간예위암세포후PTEN적표체급기재아매소유도위암세포조망중적의의.방법 (1)아매색간예위암세포BGC-823후이사갑기우담서염비색법(MTT법)화류식세포법검측세포존활솔급조망솔,병검측VFEN적mRNA화단백수평.(2)구건위암라서이위충식류,채용원위말단표기(TUNEL)법검측이위충식류중위암세포적조망정황,병용RT-PCR화Western blot법검측PTEN mRNA화단백적표체수평.(3)이PTEN특이성소간우RNA(siRNA)전염BGC-823세포,병이아매소진행간예,검측BGC-823세포적존활솔화조망솔이급PTEN단백표체수평.결과 (1)아매소간예후,BGC-823세포적생존솔정시간의뢰성강저.(2)아매소능구유효유도BGC-823세포조망.(3)아매소재BGC-823세포중가시간의뢰성지촉진PTEN적mRNA화단백수평적승고.라서이위충식류시험중,아매소간예조적조망솔[(28.11±1.05)%]명현고우대조조[(2.78±1.63)%];아매소간예조류체조직중PTEN mRNA화단백수평역고우대조조(0.5667±0.0043비0.2217±0.0063,0.14±0.26비0.04±0.15,P치균<0.05).(4)전염여미전염[WEN siRNA적위암세포이아매소간예후,PTEN siRNA전염조적PTEN단백표체수평명현저우대조조(P<0.0001),차PTEN siRNA전염조[(10.35±1.04)%]조망솔명현소우미전염조[(31.37±3.58)%],P<0.05.결론 아매소간예위암세포후가이억제기생장,유도세포조망,PTEN표체수평적승고가능시기궤제지일.
Objective To study the expression of PTEN and its significance in doxorubicin-treated gastric cancer cells. Methods (1) Gastric cancer BGC-823 cells were treated with doxorubicin. Cell proliferation and apoptosis were evaluated by MTF and flow cytometry. The expression of PTEN at the mRNA and protein level were determined by RT-PCT and Western blot, respectively. (2)The gastric cancer xenografts model was constructed. The apoptosis of gastric cancer xenografts cells was determined by TUNEL. The expression of PTEN at the mRNA and protein level were detected using RT-PCR and Western blot, respectively. (3) BGC-823 cells were transfected with PTEN siRNA before addition of doxorubicin. The proliferation and apoptosis of these cells as well as the expression level of PTEN protein were determined. Results (1) After administration of doxorubicin, the proliferation of BGC-823 cells was inhibited in a time-dependent manner. (2) Doxorubicin significantly induced apoptosis of BGC-823 cells. (3) Doxorubicin treated BGC-823 cells showed a significant increase in the expression of PTEN at the mRNA and protein level in a time-dependent manner. TUNEL assay also showed a significant increase of apoptosis rate in gastric cancer xenografts treated with doxorubicin compared with control group [(28. 11± 1.05) % vs (2. 78 ± 1.63) %]. The expression of PTEN at the mRNA and protein level in the gastric cancer xenografts were significantly increased after administration of doxorobicin (0. 5667 ± 0. 0043 vs 0.2217±0.0063,0.14±0.26 vs 0.04±0.15,P <0.05). (4) After treated with doxorubicin, the expression of PTEN in siRNA-transfected BGC-823 cells was significantly higher than that in non-transfected BGC-823 cells (P < 0. 0001). The apoptosis of PTEN siRNA-transfected BGC-823 cells was significantly decreased compared with non-transfected BGC-823 cells [(10. 35 ± 1.04) % vs (31.37 ± 3.58) %, P < 0. 05]. Conclusion Doxorubicin can effectively inhibit the growth and induce the apoptosis of BGC-823 gastric cancer cells. Increasing PTEN protein may be one of the main mechanism involved in this effect.