中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2013年
2期
229-232
,共4页
郁皓%洪婷婷%周雨峡%游庆军%吴小红%华东
鬱皓%洪婷婷%週雨峽%遊慶軍%吳小紅%華東
욱호%홍정정%주우협%유경군%오소홍%화동
唑来膦酸%胃癌%侵袭%黏附
唑來膦痠%胃癌%侵襲%黏附
서래련산%위암%침습%점부
Zoledronic acid%Gastric cancer%Invasion%Adhesion
目的 观察唑来膦酸(ZOL)对人胃癌耐阿霉素(ADM)细胞株SGC-7901/ADR侵袭和黏附的影响,探讨ZOL是否具有逆转该细胞株对ADM的耐药作用及其机制.方法 采用噻唑蓝(MTT)比色法检测1×10-4 mol/L的ZOL联合0.5 mg/L的ADM对SGC-7901/ADR细胞黏附作用的影响；Transwell实验分析上述浓度的ZOL和ADM对细胞侵袭力的影响；酶联免疫吸附试验(ELISA)法检测细胞上清液基质金属蛋白酶(MMP)-2、细胞间黏附分子-1(ICAM-1)蛋白表达水平的变化；实时定量聚合酶链反应(Real-time PCR)检测细胞MMP-2、基质金属蛋白酶抑制因子(TIMP)-2、MMP-9、ICAM-1、CD44mRNA表达水平的改变.结果 (1)1×10-4 mol/L的ZOL单药及联合0.5 mg/L的ADM与细胞共同孵育6h,黏附率分别为:对照组(C组)44.32％,ZOL单药组(Z组)34.73％,ZOL+ ADM组(Z+A组)31.30％,与C组比较,单药能够抑制细胞的黏附力(P＜0.05),Z+A组细胞黏附能力进一步下降(P＜0.01)；(2)在研究侵袭能力的实验中,C组、Z组、Z+A组每个高倍视野中细胞数(个)分别为:103.73±7.84、97.64±9.62、71.58±10.57,与对照组比较,联合用药时抑制作用显著(P＜0.05)；(3) ELISA结果显示,与C组比较、Z组及Z+A组均能够显著降低细胞上清液中的MMP-2和ICAM-1蛋白表达水平(P＜0.05或P＜0.01),与Z组比较,Z+A组差异统计学意义(P＜0.01).(4)在Real-time PCR实验中,以C组作为参考标准,药物和细胞共同孵育后,上清液CD44 mRNA表达在C组、Z组、A组、Z+A组分别为1.0000 ±0.1083、0.2601±0.0041、0.4681 ±0.0314、0.0950±0.0052,同时ICAM-1、MMP-2、MMP-9mRNA的表达在各组中也表现出类似结果.结论 ZOL对人胃癌耐ADM细胞株SGC-7901/ADR具有抑制降低细胞黏附和侵袭能力的作用,一定程度上逆转该细胞株对ADM的耐药,这些作用和下调ICAM-1、MMP-2、MMP-9、CD44 mRNA及上调TTMP-2 mRNA的表达水平相关.
目的 觀察唑來膦痠(ZOL)對人胃癌耐阿黴素(ADM)細胞株SGC-7901/ADR侵襲和黏附的影響,探討ZOL是否具有逆轉該細胞株對ADM的耐藥作用及其機製.方法 採用噻唑藍(MTT)比色法檢測1×10-4 mol/L的ZOL聯閤0.5 mg/L的ADM對SGC-7901/ADR細胞黏附作用的影響；Transwell實驗分析上述濃度的ZOL和ADM對細胞侵襲力的影響；酶聯免疫吸附試驗(ELISA)法檢測細胞上清液基質金屬蛋白酶(MMP)-2、細胞間黏附分子-1(ICAM-1)蛋白錶達水平的變化；實時定量聚閤酶鏈反應(Real-time PCR)檢測細胞MMP-2、基質金屬蛋白酶抑製因子(TIMP)-2、MMP-9、ICAM-1、CD44mRNA錶達水平的改變.結果 (1)1×10-4 mol/L的ZOL單藥及聯閤0.5 mg/L的ADM與細胞共同孵育6h,黏附率分彆為:對照組(C組)44.32％,ZOL單藥組(Z組)34.73％,ZOL+ ADM組(Z+A組)31.30％,與C組比較,單藥能夠抑製細胞的黏附力(P＜0.05),Z+A組細胞黏附能力進一步下降(P＜0.01)；(2)在研究侵襲能力的實驗中,C組、Z組、Z+A組每箇高倍視野中細胞數(箇)分彆為:103.73±7.84、97.64±9.62、71.58±10.57,與對照組比較,聯閤用藥時抑製作用顯著(P＜0.05)；(3) ELISA結果顯示,與C組比較、Z組及Z+A組均能夠顯著降低細胞上清液中的MMP-2和ICAM-1蛋白錶達水平(P＜0.05或P＜0.01),與Z組比較,Z+A組差異統計學意義(P＜0.01).(4)在Real-time PCR實驗中,以C組作為參攷標準,藥物和細胞共同孵育後,上清液CD44 mRNA錶達在C組、Z組、A組、Z+A組分彆為1.0000 ±0.1083、0.2601±0.0041、0.4681 ±0.0314、0.0950±0.0052,同時ICAM-1、MMP-2、MMP-9mRNA的錶達在各組中也錶現齣類似結果.結論 ZOL對人胃癌耐ADM細胞株SGC-7901/ADR具有抑製降低細胞黏附和侵襲能力的作用,一定程度上逆轉該細胞株對ADM的耐藥,這些作用和下調ICAM-1、MMP-2、MMP-9、CD44 mRNA及上調TTMP-2 mRNA的錶達水平相關.
목적 관찰서래련산(ZOL)대인위암내아매소(ADM)세포주SGC-7901/ADR침습화점부적영향,탐토ZOL시부구유역전해세포주대ADM적내약작용급기궤제.방법 채용새서람(MTT)비색법검측1×10-4 mol/L적ZOL연합0.5 mg/L적ADM대SGC-7901/ADR세포점부작용적영향；Transwell실험분석상술농도적ZOL화ADM대세포침습력적영향；매련면역흡부시험(ELISA)법검측세포상청액기질금속단백매(MMP)-2、세포간점부분자-1(ICAM-1)단백표체수평적변화；실시정량취합매련반응(Real-time PCR)검측세포MMP-2、기질금속단백매억제인자(TIMP)-2、MMP-9、ICAM-1、CD44mRNA표체수평적개변.결과 (1)1×10-4 mol/L적ZOL단약급연합0.5 mg/L적ADM여세포공동부육6h,점부솔분별위:대조조(C조)44.32％,ZOL단약조(Z조)34.73％,ZOL+ ADM조(Z+A조)31.30％,여C조비교,단약능구억제세포적점부력(P＜0.05),Z+A조세포점부능력진일보하강(P＜0.01)；(2)재연구침습능력적실험중,C조、Z조、Z+A조매개고배시야중세포수(개)분별위:103.73±7.84、97.64±9.62、71.58±10.57,여대조조비교,연합용약시억제작용현저(P＜0.05)；(3) ELISA결과현시,여C조비교、Z조급Z+A조균능구현저강저세포상청액중적MMP-2화ICAM-1단백표체수평(P＜0.05혹P＜0.01),여Z조비교,Z+A조차이통계학의의(P＜0.01).(4)재Real-time PCR실험중,이C조작위삼고표준,약물화세포공동부육후,상청액CD44 mRNA표체재C조、Z조、A조、Z+A조분별위1.0000 ±0.1083、0.2601±0.0041、0.4681 ±0.0314、0.0950±0.0052,동시ICAM-1、MMP-2、MMP-9mRNA적표체재각조중야표현출유사결과.결론 ZOL대인위암내ADM세포주SGC-7901/ADR구유억제강저세포점부화침습능력적작용,일정정도상역전해세포주대ADM적내약,저사작용화하조ICAM-1、MMP-2、MMP-9、CD44 mRNA급상조TTMP-2 mRNA적표체수평상관.
Objective To detect effects of zoledronic acid on the invasion and adhesion of adriamycin resistant gastric cancer cell line SGC-7901/ADR,and to investigate whether zoledronic acid could restore sensitivity to adriamycin and possible mechanism.Methods Methyl thiazol tetrazolium (MTT) assay was used to investigate the effect of 1 × 10-4 mol/L zoledronic acid and 0.5 mg/L doxorubicin on the adhesion of SGC7901/ADR cells.Transwell was used to investigate cellular invasive ability.Enzyme linked immunosorbent assay (ELISA) was used to quantify supematant matrix metalloproteinase (MMP)-2 and intercellular adhesion molecule-1 (ICAM-1).MMP-2,tissue inhibitorof metalloproteinase (TIMP)-2,MMP-9,ICAM-1 and CD44 levels were quantified by using real-time polymerase chain reaction (PCR).Results (1) After SGC-7901/ADR cells were incubated with 1 × 10-4 mol/L zoledronic acid (Z) alone or in combined with 0.5 mg/L adriamycin (Z + A) for 6 h,the adhesion rate was 44.32％ for control (C)group,34.73％ for Z group,and 31.30％ for Z + A group.The adhesion rate was reduced in treated groups as compared with untreated group (P ＜ 0.05) ; (2) The number of invasive cells per high power field in C group,Z group and Z + A group was 103.73 ±7.84,97.64 ±9.62,and 71.58 ± 10.57 respectively.As compared with control group,the invasive ability was reduced significantly in Z + A group (P ＜ 0.05) ; (3) ELISA results indicated that as compared with control group,the MMP-2 and ICAM-1 in the supernatant of Z group and Z + A group were significantly reduced (P ＜0.05 or P ＜0.01).As compared with Z group,both MMP-2 and ICAM-1 were significantly reduced in Z + A group (P＜0.01) ;(4) CD44 mRNA expression in C group,Z group,and Z +A group was 1.0000 ±0.1083,0.2601 ±0.0041,0.4681 ±0.0314,and 0.0950 ± 0.0052 respectively.Similar results were observed in the expression of ICAM-1,MMP-2 and MMP-9.Conclusion Zoledronic acid could inhibit the adhesion and invasion of SGC-7901/ADR cells,and restore the sensitivity of SGC-7901/ADR cells to doxorubicin.These effects may be related to the decreased expression of ICAM-1,MMP-2,MMP-9 and CD44,and the increased TIMP-2 expression.
