增韧基团-姜黄素酯协同二甲双胍对前列腺癌PC-3细胞凋亡的影响

Synergistic effect of toughening perssad-curcumin with ester and metformin on apoptosis of andro-genic independent prostate cancer cell line PC-3

目的探讨姜黄素酯协同二甲双胍抑制雄激素非依赖性前列腺癌细胞株PC-3增殖的影响及其机制。方法细胞免疫组织化学法初步鉴定原代与传代PC-3细胞中CD133、CD44的表达。3种浓度的叔丁氧羰基一苯丙氨酸酯姜黄素单酯（BPC）联合二甲双胍作用于人前列腺癌Pc-3细胞，与单独使用作比较。采用噻唑蓝（MTT）比色法检测上述10组细胞生长活性，流式细胞术检测细胞凋亡和比率。结果原代PC-3细胞CD44、CDl33阳性表达强度（56．5±1．0）％、（50．7±4．0）％明高显于传代Pc-3细胞（23．1±1．1）％、（18．5±2．5）％（P〈0．05）；二甲双胍（5mmol／L）组、不同浓度BPC（10、20、40μmol／L）组、二甲双胍（5mmol／L）联合不同浓度BPC组分别作用原代PC-3细胞24h后细胞生长抑制率分别为（20．3±1．7）％、（15．3±1．1）％～（49．3±2．0）％、（50．9±1．0）％-（68．4±3．2）％；二甲双胍（5mmol／L）、BPC40μmol／L、二甲双胍＋BPC（5mmol／L，40μmol／L）作用于PC-3细胞24h后的细胞凋亡率为（21．02±2．10）％、（28．04±1．40）％、（45．03±2．80）％，联合用药组与相应对照组间细胞抑制率及凋亡率差异有统计学意义（P〈0．05）。二甲双胍（5mmol／L）作用于传代前列腺癌Pc-3细胞24h后细胞增殖抑制率为（4．95±1．20）％，细胞凋亡率为（5．02±1．70）％，较原代细胞抑制率及凋亡率均降低差异有统计学意义（P〈0．05）。结论增韧基团姜黄素酯可与二甲双胍协同抑制激素非依赖性前列腺癌Pc-3细胞的增殖。其协同机制可能为：增韧基团姜黄素酯靶向杀伤前列腺癌PC-3细胞中的癌细胞，而二甲双胍靶向杀伤前列腺癌PC-3细胞中的肿瘤干细胞。

Objective To study the effect of toughening genes-linking curcumin with ester and Metforrnin on apoptosis of androgenic independent prostate cancer cell line PC-3. Methods Immunohisto- chemical staining was used to detect the distribution and expression intensity of CD44 and CD133 antigens in PC-3 cells of primary and 8th passage. PC-3 ceils were treated with three concentrations of Boc-pheny- lalanine-curcumin （BPC） combined with Metformin, BPC or Metformin alone respectively for 24 h. The growth activities of cancer cells were studied by methyl thiazol tetrazolium （MTT） colorimetry. Cell apopto- sis and its rate were detected by using flowcy-tometry. The morphological changes of cancer cells were ob- served under electronic microscopy. Results The positive expression rate of CIM4 and CD133 antigens in primary PC-3 cells was （56. 5±1.0） % and （50. 7±4. 0） % respectively, which was significantly higher than that in passaged PC-3 cells [ （23.1±1.1 ）% and （ 18.5 ±2. 5% ） respectively]. After treatment with 5 mmol/L Metformin, 10-40 μmol/L BPC, 5 mmol/L Metformin ＋ 10-40 μmol/L BPC for 24 h, the growth inhibition rate of PC-3 cells was （20.3±1.7）%, （ 15.3 ±1.1 ）%-（49.3± 2.0）%, and （50. 9±1.0）%-（ 68.4 -±3.2 ）% respectively. After treatment with 5 mmol/L Metformin, 40 Ixmol/L BPC, 5 mmol/LMetformin plus 40 μmol/L BPC for 24 h, the apoptosis rate was （21.02±2. 10 ） %, （28.04±1.40）% , and （45.03 ＋2. 80）% respectively. There was significant difference in the growth in- hibition rate and the apoptosis rate between Metformin plus BPC group and corresponding control group （P 〈0. 05）. After treatment with 5 mmol/L Metformin, the growth inhibition and apoptosis rate of pas- saged PC-3 cells was （4.95±1.20）% and （5.02± 1.70）% respectively, which was significantly lower than that in primary PC-3 cells. Conclusion Metformin can enfi）rce antiproliferative effect of BPC aganist human prostatic cancer eell line PC-3. Tumor stem cells may exist in prostate cancer PC-3 cells. Metformin possiby targetedly kills tumor stem cells which exist in prostate cancer PC-3 cells.