Increased sensitivity of human lung adenocarcinoma cells to cisplatin associated with downregulated contactin-1

Contactin-1 (CNTN-1), a glycosyl phosphatidylinositol anchor neural cell adhesion molecule (ACAM), is thought to function not only in nervous system development but also in the invasion and metastasis of several tumours. To investigate whether CNTN-1 is involved in multidrug resistance (MDR) in lung adenocarcinoma, CNTN-1 expression was compared between MDR human lung adenocarcinoma A549/cisplatin (A549/DDP) cells and its progenitor A549 cells. The comparison showed that CNTN-1 expression in A549/DDP cells was significantly higher than in A549 cells both at the mRNA level and the protein level. In order to confirm the physiological function of the abnormal expression, lentivirus-mediated short hairpin RNA (shRNA) was used to silence CNTN-1. Cell cytotoxicity assay and cell apoptosis assay revealed that silencing CNTN-1 both in A549 cells and in A549/DDP cells not only rendered cells more sensitive to cisplatin than the negative control, but also increased the cisplatin-induced apoptosis. Metastasis and invasion assays demonstrated that CNTN-1 knockdown reduced metastasis and invasion but did not affect A549 or A549/DDP cell proliferation. To investigate whether the abnormal expression of CNTN-1 is associated with characteristics of patients with non-small cell lung cancer (NSCLC), immunohistochemistry was used to detect CNTN-1 expression in 143 tissue samples from NSCLC patients and the results showed that the degree of CNTN-1 expression positively correlated with lymphatic invasion in patients with lung adenocarcinoma who received adjuvant cisplatin- or carboplatin-based treatment after surgery. Thus, we concluded that CNTN-1 is closely related with MDR of lung adenocarcinoma. Additionally, CNTN-1 is a novel marker to predict chemotherapeutic efficacy of patients with lung adenocarcinoma, especially with regard to cisplatin- or carboplatin-based regimens.     

肿瘤坏死因子α对人肺腺癌细胞耐顺铂的影响

目的研究肿瘤坏死因子OL（TNF-α）对人肺腺癌细胞系A549／顺铂（DDP）耐DDP的逆转作用,探讨其与肺耐药相关蛋白（LRP）表达之间的关系。方法以MTT法检0n,0TNF-α与顺铂联用对A549／DDP细胞的细胞毒性作用,以免疫细胞化学方法检测A549／DDP细胞LRP的表达情况。结果250、1000U／mlTNF-α可使顺铂对A549／DDP细胞的IC50从7．12ng／L分别降至5．02、4．41ng／L,能够逆转A549／DDP对顺铂的耐药,逆转倍数分别为1．42、1．62。A549／DDP细胞LRP呈强阳性表达,250、1000U／mlTN-α与顺铂联用可以下调LRP表达,LRP阳性表达率分别为（60．14-4-6．54）％、（57．234-5．98）％,同无药组和顺铂组LRP表达率（79．63±4．78）％、（75．97±5．32）％比较,差异具有统计学意义（P均〈0．01）。结论TNF-α能够逆转A549／DDP对顺铂的耐药,其机制可能与下调LRP表达有关。     

Super enhancer‐LncRNA SENCR promoted cisplatin resistance and growth of NSCLC through upregulating FLI1

BackgroundSuper enhancer‐lncRNA smooth muscle and endothelial cell‐enriched migration/differentiation‐associated lncRNA (SENCR) were highly overexpressed in cisplatin‐resistant A549/DDP cells, while the mechanism was unclear.MethodsSE‐lncRNA SENCR and FLI1 mRNA expression in A549/DDP cell, LAD tissues were detected. SENCR knockdown of A549/DDP cell and SENCR overexpression of cisplatin‐sensitive A549 cell were constructed. Experiments of cell‐confirmed function of SENCR and the correlation between SENCR and FLI1 were validated.ResultsThe expression of SENCR and FLI1 mRNA in A549/DDP cell were both upregulated and mainly localized in the nucleus. Compared with DDP‐sensitive tissues with disease relief, SENCR expression was higher in DDP‐resistant tissues with disease progression from LAD. Knockdown of SENCR in A549/DDP reduced proliferation ability and cisplatin resistance, consistent with the decreased levels of proteins PCNA, MDMX, and P‐gp. Besides, whatever without cisplatin or with 2 μg/ml cisplatin, knockdown of SENCR reduced the migration, invasion, and colony formation abilities of A549/DDP cell and promoted apoptosis. However, when SENCR was overexpressed in A549 cell, all above results were reversed. Mechanistically, FLI1 expression was reduced after knocking down SENCR, while overexpressing SENCR increased FLI1 expression.ConclusionSE‐LncRNA SENCR was upregulated in A549/DDP, which could promote cisplatin resistance and growth of NSCLC cell through upregulating FLI1 expression.     

RNA干扰ERCC1基因对肺癌细胞药物敏感性的影响

目的 探讨RNA干扰切除修复交叉互补基因1(ERCC1)对肺癌细胞药物敏感性的影响.方法 将肺癌A549/DDP细胞分为阴性组、空白组、ERCC1-shRNA1组及ERCC1-shRNA2组,通过不同方法检测分析ERCC1的mRNA和蛋白表达水平,及不同顺铂(DDP)浓度下肺癌细胞的凋亡程度.结果 阴性组、空白组的ER...     

mir-17-5p、mir-92a、let-7b表达水平与非小细胞肺癌顺铂耐药的关系

目的探讨mir-17-5p、mir-92a、let-7b表达水平与非小细胞肺癌顺铂耐药关系。方法以人非小细胞肺癌细胞系A549及其耐药株A549/DDP为研究对象,采用RT-PCR法检测mir-17-5p、mir-92a及let-7b在细胞中的表达水平,采用cck8检测其细胞存活情况,采用细胞克隆平台方法,检测转染前后细胞的增殖情况,采用流式细胞仪检测转染前后细胞的凋亡情况。结果（1） A549/DDP细胞mir-17-5p的表达水平是A549细胞的2.11±0.25倍（P＜0.05）;A549/DDP细胞mir-92a的表达水平是A549细胞的 7.40 ± 1.05 倍（P＜0.05）;而A549/DDP 细胞let-7b 的表达水平是A549 细胞的（26.54 ± 2.90）%（P＜0.05）;（2）A549 转染mir-17-5pmimic,mir-92a mimic 以及let-7b inhibitor 后对顺铂的敏感性下降（P＜0.05）;A549/ddp 转染mir-17-5p inhibitor, mir-92a inhibitor以及let-7b mimic后对顺铂的敏感性增加（P＜0.05）;（3）A549转染mir-17-5p mimic,mir-92a mimic以及let-7b inhibitor 后,细胞形成克隆集落数量数量多于对照组（P＜0.05）;而A549/ddp 转染mir-17-5p inhibitor,mir-92a inhibitor 以及 let-7b mimic 后,细胞形成克隆集落数量数量少于对照组（P＜0.05）;（4）A549 转染mir-17-5p mimic,mir-92a mimic 以及let-7b inhibitor后,细胞凋亡率明显低于对照组（P＜0.05）;而a549/ddp转染mir-17-5p inhibitor,mir-92a inhibitor以及let-7b mimic后, 细胞凋亡率明显高于对照组（P＜0.05）。结论Mir-17-5p、mir-92a表达水平升高,let-7b表达水平下降,可以促进肺癌细胞增殖, 抑制其凋亡以及使肺癌细胞对顺铂敏感性下降。     

肺腺癌细胞A549及其顺铂耐药细胞株A549/DDP中GSK-3β蛋白磷酸化水平的差异

目的研究人肺腺癌细胞系A549和其顺铂耐药细胞系A549/DDP中GSK-3β的磷酸化和胞内分布以探讨GSK-3β在顺铂耐药中的作用。方法蛋白质免疫印迹法检测A549/DDP和A549细胞质和细胞核中总GSK-3β、p-GSK-3βser9和p-GSK-3βtyr6的表达。MTT法、流式细胞术分别检测顺铂耐药性、肺癌细胞凋亡率。结果 A549/DDP细胞胞质中p-GSK-3βser9水平明显高于A549细胞（P〈0.01）,顺铂的处理增加了A549/DDP细胞中p-GSK-3βser9的水平（P〈0.01）,相反却减少了A549细胞中p-GSK-3βser9的水平（P〈0.01）。A549/DDP细胞质中p-GSK-3βtyr6水平明显低于A549细胞（P〈0.01）,顺铂的处理减少了A549/DDP细胞中p-GSK-3βtyr6的水平（P〈0.01）,却增加了A549细胞中p-GSK-3βtyr6的水平（P〈0.01）。结论细胞质中GSK-3β活性受抑可能是非小细胞肺癌顺铂耐药的原因。     

miR-802对叉头框转录因子M1抑制A549/DDP细胞顺铂耐药性的靶向调控作用

目的探讨miR-802抑制非小细胞肺癌A549/DDP细胞的顺铂耐药性及其对叉头框转录因子M1(forkhead box protein M1, FoxM1)的靶向调控作用。方法 A549、A549/DDP细胞采用反转录PCR法检测miR-802相对表达量。将A549/DDP细胞分为空白转染组和miR-802过表达组(过表达组),分别转染miR-NC和miR-802类似物(miR-802 mimics),转染24、48、72、96 h后,采用反转录PCR法检测2组细胞miR-802相对表达量,并采用CCK-8法检测2组细胞增殖率;转染48 h后,CCK-8法检测转染后A549/DDP细胞对DDP的敏感性,采用流式细胞仪检测2组细胞凋亡率和细胞周期,采用Western blot法检测转染后A549/DDP细胞内FoxM1蛋白相对表达量。结果 A549/DDP细胞miR-802相对表达量(0.21±0.03)低于A549细胞(0.85±0.12)(P<0.05);转染24、48、72、96 h,过表达组细胞miR-802相对表达量依次增高(P<0.05),且均高于空白转染组(P<0.05);转染48、72、96 h,空白转染组细胞增殖率依次增高(P<0.05),且均高于过表达组(P<0.05);转染48 h,过表达组细胞半数抑制浓度[(35.28±2.17)mg/L]和细胞早期凋亡率[(17.2±1.1)%]均高于空白转染组[(14.22±1.28)mg/L、(9.0±0.8)%](P<0.05),S期和G2/M期细胞比率[(21.30±0.20)%、(8.35±0.33)%]及细胞内FoxM1蛋白相对表达量(0.21±0.04)均低于空白转染组[(27.54±0.52)%、(14.67±0.70)%、0.44±0.06](P<0.05)。结论 miR-802可能通过抑制FoxM1表达而降低非小细胞肺癌A549/DDP细胞的顺铂耐药性。     

紫杉醇耐药人肺腺癌细胞系的建立及生物学活性

目的：体外建立紫杉醇耐药人肺腺癌A549/TAX细胞系,并对其生物学特性进行研究.方法：采用逐步增加紫杉醇浓度结合低剂量持续诱导法,建立紫杉醇耐药人肺腺癌A549/TAX细胞系.MT T法描绘A549和A549/TAX细胞的生长曲线;流式细胞仪检测细胞周期并比较经紫杉醇诱导24 h后A549和A549/TAX细胞平均凋亡率的差异;检测紫杉醇对A549/TAX细胞的半数抑制浓度、耐药系数（RI）及A549/TAX细胞对其他 5种化疗药物（顺铂、长春新碱、表阿霉素、足叶乙甙及吉西他滨）的交叉耐药谱;RT-PCR半定量分析A549/TAX细胞凋亡基因（bcl-2）及耐药基因（MDR1、LRP、GST-π）的mRNA表达.结果：A549/TAX细胞生长较亲本细胞快,S期细胞增多[（51.61±0.48）%],G1 期减少[（37.26±0.23）%],经紫杉醇（150 μmol/L）诱导前后平均凋亡率差异与A549细胞相比差异有显著性.A549/TAX细胞对紫杉醇、长春新碱、表阿霉素及足叶乙甙的RI分别为 21.3、12.91、5.88和4.79,而对顺铂（RI=1.06）和吉西他滨（RI=1.03）则无交叉耐药;A549/TAX细胞bcl-2、 MDR1、LRP mRNA的表达均较亲本细胞显著增高（P＜0.001）,GST-πmRNA未见表达.结论：成功建立了紫杉醇耐药A549/TAX细胞系,该细胞抗药性能明显、稳定,推测其多药耐药性与bcl-2、MDR1和LRP高表达有关.     

热化疗联合对人肺腺癌耐药细胞株A549／DDP作用及其机制的实验研究

目的 观察热化疗联合对人肺腺癌耐药细胞株A549／DDP的影响及其机制，为临床上应用热化疗联合治疗非小细胞肺癌提供理论依据。方法 CCK-8法检测A549、不同温度下A549／DDP细胞对顺铂的敏感性；流式细胞仪分析热疗对A549／DDP细胞周期分布的影响；RT-PCR检测热疗对A549／DDP细胞ERCC1表达的影响。结果 亲代人肺腺癌细胞株A549对顺铂的敏感性明显高于耐药细胞株ASg9／DDP，不同温度加热后A549／DDP细胞对顺铂的敏感性均有提高；热疗尤其是热化疗后G2／M期细胞明显增多，G0／G1期细胞明显减少（P〈0．01）；热疗后A549／DDP细胞ERCC1mRNA表达未见明显改变。结论 热疗能提高人肺腺癌耐药细胞株A549／DDP对顺铂的敏感性，热化疗具有协同作用。热化疗协同作用机制可能主要是诱导细胞周期阻滞，抑制细胞增殖，和DNA修复基因ERCC无直接相关性。     

HIF-2α基因与人肺腺癌A549细胞株对顺铂耐药性的关系

目的：探讨小干扰RNA（small interfering RNA,siRNA）沉默缺氧诱导因子2α（hypoxia inducible factor-2 alpha,HIF2α）基因表达后,人肺腺癌A549细胞株对顺铂耐药性的变化.方法：构建靶向HIF-2α基因的siRNA真核表达载体并转染A549细胞,采用Reαl-time PCR检测HIF-2α mRNA的表达,采用Western blotting检测HIF-2α及多药耐药基因1（multidrug resistance-1,Mdr-1）蛋白的表达,采用MTT法检测顺铂处理后细胞的存活率.结果：在HIF-2α siRNA转染A549细胞后24、48 h后,HIF-2α在mRNA和蛋白水平的表达与空白对照组相比均显著下调（均P＜0.01）,Mdr 1蛋白的表达水平也明显降低（均P＜0.05）.与空白对照组相比,HIF-2α siRNA组细胞对顺铂的敏感性明显增加,各时间点的IC50值均显著降低,差异有统计学意义（均P＜0.01）.结论：靶向HIF-2α的siRNA表达载体能够有效抑制HIF-2α基因的表达,从而逆转A549细胞对顺铂的化疗耐药性.     

DNA修复相关基因甲基化与肺癌A549细胞顺铂化疗耐药的相关性研究

目的探讨5-氮杂-2’脱氧胞苷（5-Aza-2’deoxycytidine,5-Aza—Cde）对顺铂耐药株肺癌A549／DDP细胞DNA修复相关基因hMLH1、MGMT基因启动子区DNA甲基化状态及其表达的影响。方法A549细胞（A549细胞组）、A549／DDP细胞（A549／DDP组）、A549／DDP细胞5、10、20μmol／L5-Aza—Cde组,分别采用甲基化特异性PCR检测细胞hMLHl、MGMT基因甲基化状态,反转录一PCR法检测用药前、后细胞hMLHlmRNA、MGMTmRNA表达情况。结果A549细胞组hMLHl基因呈非甲基化状态且高表达,MGMT呈部分甲基化状态且高表达,A549一DDP细胞组hMLHl、MGMT基因均呈高甲基化状态,mRNA表达下调,10、20μmol／L5-Aza—Cde组hMLHl和MGMT均呈非甲基化状态;A549／DDP细胞组、5μmol／L5-Aza—Cde组hMLHlmRNA与MGMTmRNA表达量均低于A549细胞组（P〈0．05）,20μmol／L5-Aza—Cde组与A549细胞组比较差异无统计学意义（P〉0．05）;10μmol／L5-Aza—Cde组MGMTmRNA表达量低于A549细胞组（P〈0．05）,hMLH1mRNA表达量与A549细胞组比较差异无统计学意义（P〉0．05）。结论hMLHl、MGMT基因甲基化修饰程度与mRNA表达有一定相关性,hMLH1、MGMT基因甲基化可能是肺癌A549细胞对顺铂耐药的因素之一。     

抑制葡萄糖调节蛋白78基因表达逆转卵巢癌顺铂耐药作用的实验研究

目的探讨以葡萄糖调节蛋白78(GRP78)基因为靶向的RNA干扰(RNAi)抑制GRP78基因的表达,对人卵巢癌顺铂耐药细胞株SKOV3/DDP的影响。方法以GRP78基因为靶向的pSilencerTM3.0-H1-GRP78 siRNA重组质粒的构建,脂质法转染至细胞中;RT-PCR及Western blot方法检测GRP78 mRNA及蛋白表达;Western blot方法检测CHOP蛋白表达;MTT法检测不同浓度顺铂作用下细胞存活率,计算IC50及耐药指数。结果 RT-PCR及Western blot检测转染GRP78 siRNA重组质粒24 h、48 h7、2 h均能明显抑制SKOV3/DDP细胞GRP78基因表达,其中48h抑制效果最为明显;抑制SKOV3/DDP细胞GRP78基因表达能上调CHOP蛋白表达;顺铂作用24h,SKOV3/DDP细胞的IC50(26.13μg/ml)是其亲本SKOV3细胞IC50(8.25μg/ml)的3.1倍;转染pSH1Si-GRP78重组质粒后SKOV3/DDP细胞IC50(11.62μg/ml)明显降低。结论转染GRP78 siRNA重组质粒有效抑制SKOV3/DDP细胞GRP78基因表达;抑制GRP78基因表达能逆转SKOV3/DDP细胞的顺铂耐药性。     

小分子Survivin干扰RNA逆转肺癌细胞耐药的研究

目的探讨小分子Survivin干扰RNA（siRNA-Survivin）逆转人肺癌耐药细胞（A549DDP）耐药性的研究。方法应用RT-PCR法及Western-blot方法证明Survivin基因在人肺腺癌耐药细胞系（A549DDP）细胞系呈现高表达;并采用脂质体为转染介质,将Survivin小片段RNA导入A549DDP细胞沉默Survivin基因;应用MTT法观察顺铂、多西他赛、吉西他滨、表阿霉素对沉默Survivin基因后的A549DDP细胞的细胞增殖抑制及半数抑制浓度的变化。结果 1,Sur-vivin-mRNA、Survivin蛋白在A549DDP细胞系表达水平均显著高于A549细胞系;2,顺铂、多西他赛、吉西他滨、表阿霉素对沉默Survivin后A549DDP细胞的细胞增殖抑制与对照组（A549DDP细胞s、iRNA-control A549DDP细胞）相比明显增加,半数抑制浓度（IC50）明显下降,差异显著（P〈0.01）。结论 Survivin参与了肺腺癌细胞耐药的形成;小分子Sur-vivin干扰RNA沉默Survivin可以提高耐药的肺癌细胞对化疗药物的敏感性。Survivin可能成为临床监测肿瘤耐药性变化,逆转肺癌耐药的一个新的靶点。     

Soluble CD40 ligands sensitize the epithelial ovarian cancer cells to cisplatin treatment

ObjectiveCD154 (CD40L) is a protein that is primarily expressed on activated T cells and is a member of the TNF superfamily of molecules. It binds to CD40 on antigen-presenting cells (APC), which leads to many effects depending on the target cell type. Being an activator of immune cells, CD40L has also been shown to directly induce apoptosis in tumor cells by multiple mechanisms. To understand the role of sCD40L in regulating the proliferation of epithelial ovarian cancer cells treated or untreated with cisplatin.MethodsEpithelial ovarian cancer cells: SKOV3 and its cisplatin-resisitant strain SKOV3/DDP cells were used to test the effect of sCD40L and cisplatin. The proliferation of SKOV3 and SKOV3/DDP cells were measured by MTT. Cell cycle was assessed by flow cytometry. The mRNA expressions of targeted genes were detected by qRT-PCR. The protein expressions were detected by Western blotting.ResultssCD40L showed a significant dose-dependence inhibitory effect on the proliferation of ovarian cancer cell lines. sCD40L in combination with cisplatin could sensitized SKOV3/DDP cells to cisplatin treatment and reversed the drug resistance of SKOV3/DDP cells. The reversal ratios of 1 μg/ml sCD40L combined with cisplatin in SKOV3 and SKOV3/DDP cells were 2.11, 2.71, while the reversal ratios of 2 μg/ml sCD40L combined with cisplatin in SKOV3 and SKOV3/DDP cells were 3.78, 5.20, respectively. sCD40L or sCD40L combined cisplatin increased tumor cells in G0/G1 phase. sCD40L in combination with cisplatin decreased the expression levels of GST-π, LRP, Survivin, p53 and Bcl-2 in both epithelial ovarian cancer cell lines. The protein expression level of GST-π, LRP and P53 protein was also decreased upon sCD40L in combination with cisplatin although the expression level of Bcl-2 and survivin protein had no significant difference.ConclusionsCD40L inhibits the proliferation of SKOV3 and SKOV3/DDP cells. The combined application of sCD40L and cisplatin can strength the inhibitory effect of cisplatin, and to a certain extent, reversing the resistance to cisplatin in SKOV3/DDP cells. sCD40L could lead a cell block in G0/G1 phase and make the cell growth restrained. sCD40L could induce SKOV3 and SKOV3/DDP cells apoptosis and reverse drug resistance through cutting GST-π mRNA, LRP mRNA, survivin mRNA, p53 mRNA and Bcl-2 mRNA and decreasing the expression of GST-π, LRP and P53 protein in SKOV3 and SKOV3/DDP cells, which provides in-vivo experiment basis to the application of sCD40L as a drug improving ovarian cancer cells sensitivity to cisplatin.     

奈达铂与顺铂对膀胱癌细胞系BIU-87作用的体内外实验比较

目的比较奈达铂与顺铂对膀胱癌细胞系BIU-87的体内外杀伤作用及毒副作用。方法用MTT法检测比较奈达铂与顺铂对BIU-87细胞系的抑制率;用裸鼠接种实验检测比较奈达铂与顺铂对裸鼠BIU-87移植瘤的抑制作用及毒性作用。结果奈达铂在体外对BIU-87细胞系的抑制率与顺铂相似,但对裸鼠移植瘤抑制力更强。奈达铂毒性反应较轻。结论在膀胱癌化疗中,奈达铂可能比顺铂更具优势。     

RNA干扰核糖核苷酸还原酶M2对耐药卵巢癌细胞凋亡及侵袭性的影响

目的探讨RNA干扰核糖核苷酸还原酶M2(RRM2)对耐药卵巢癌细胞凋亡及侵袭性的影响。方法将RRM2基因的特异性小干扰(siRNA)转染SKOV3/DDP设为干扰组,SKOV3/DDP细胞、SKOV3/DDP-RRM2非特异性阴性细胞设为空白组、阴性组。检测细胞增殖抑制率,计算RI、耐药转染率,荧光PCR技术检测RRM2基因mRNA表达,并分析siRNA转染对SKOV3/DDP耐药指数、RRM2蛋白的影响,采用Transwell观察SKOV3/DDP细胞侵袭能力。结果空白组、阴性组及干扰组转染率均90%。DDP对SKOV3/DDP细胞属低度耐药,吉西他滨对SKOV3/DDP细胞仍较为敏感。DDP、吉西他滨对干扰组、阴性组、空白组的DDP对细胞的半数抑制浓度(IC50)值比较,差异有统计学意义(P0.05),DDP、吉西他滨对干扰组细胞的IC50值显著低于阴性组、空白组(P0.05)。SKOV3/DDP细胞、SKOV3细胞RRM2蛋白的相对表达量比较,差异有统计学意义(P0.05),且转染Ⅰ组、Ⅱ组、Ⅲ组相对表达量均显著低于阴性组、空白组,其中以转染Ⅰ组细胞的RRM2蛋白相对表达量下降最显著(P0.05)。干扰组细胞凋亡率显著高于空白组、阴性组,穿膜细胞数显著低于空白组、阴性组(P0.05)。结论 siRNA可有效抑制RRM2基因在卵巢癌中的增殖与侵袭性,增加耐药细胞的药物敏感性,尤其是促进DDP诱导的耐药细胞凋亡。     

MiR-520a对胃癌SGC7901细胞增殖、侵袭、迁移以及顺铂药物敏感性的影响

目的:探讨miR-520a在胃癌中的表达以及对胃癌细胞生物学功能的影响。方法:通过real-time PCR检测miR-520a在胃癌组织和胃癌细胞中的表达。采用慢病毒在胃癌细胞SGC7901中过表达miR-520a,在体外观察miR-520a对胃癌细胞增殖、凋亡、侵袭和迁移以及对顺铂(DDP)敏感性的影响。并通过异种移植瘤实验观察过表达miR-520a对胃癌细胞在裸鼠体内生长的影响。结果:与癌旁正常组织相比,miR-520a在胃癌组织中的表达明显降低。与正常胃黏膜上皮细胞GES-1相比,miR-520a在胃癌SGC7901细胞中的表达明显降低,且在顺铂耐药SGC7901/DDP细胞中的表达更低。miR-520a过表达能够促进SGC7901细胞凋亡,抑制细胞增殖、侵袭和迁移。miR-520a过表达促进SGC7901细胞对DDP的药物敏感性增加。异种移植瘤实验发现miR-520a过表达的细胞在裸鼠体内形成的肿瘤生长速度减慢。结论:miR-520a可抑制胃癌发生与发展,提高胃癌细胞对顺铂的敏感性。     

姜黄素联合顺铂对肺癌治疗的体外实验研究

目的探讨姜黄素(CUR)与化疗药(DDP)联合应用治疗肺癌的效应和可能机制。方法应用MTT试验检测CUR与DDP联用对人肺癌A549细胞增殖的影响,用流式细胞术检测CUR与DDP联用对人肺癌A549细胞周期和凋亡的影响。结果在一定的浓度范围内,随着姜黄素与顺铂均可抑制细胞生长,呈量-效关系。姜黄素与顺铂联用时,可以增强对A549细胞的增殖抑制作用。姜黄素和顺铂均可诱导A549细胞的凋亡,而且两者联用可增加A549细胞的凋亡率。姜黄素将细胞聚结在G2/M期并可诱导凋亡,顺铂将细胞聚结在S期并可诱导凋亡。结论姜黄素、顺铂均可抑制人肺腺癌A-549细胞的增殖、诱导细胞的凋亡,在一定浓度范围内,呈量-效关系,而且两者联合应用具有相加或协同作用。其效应可能是通过对细胞周期的影响来实现的。     

姜黄素联合顺铂对肺癌治疗的体外实验研究

目的探讨姜黄素（CUR）与化疗药（DDP）联合应用治疗肺癌的效应和可能机制。方法应用MTT试验检测CUR与DDP联用对人肺癌A549细胞增殖的影响，用流式细胞术检测CUR与DDP联用对人肺癌A549细胞周期和凋亡的影响。结果在一定的浓度范围内，随着姜黄素与顺铂均可抑制细胞生长，呈量-效关系。姜黄素与顺铂联用时，可以增强对A549细胞的增殖抑制作用。姜黄素和顺铂均可诱导A549细胞的凋亡，而且两者联用可增加A549细胞的凋亡率。姜黄素将细胞聚结在C2／M期并可诱导凋亡，顺铂将细胞聚结在s期并可诱导凋亡。结论姜黄素、顺铂均可抑制人肺腺癌A-549细胞的增殖、诱导细胞的凋亡，在一定浓度范围内，呈量-效关系，而且两者联合应用具有相加或协同作用。其效应可能是通过对细胞周期的影响来实现的。     

肿瘤坏死因子相关凋亡诱导配体对卵巢癌裸鼠移植瘤SKOV3细胞的促凋亡研究

目的 研究肿瘤坏死因子相关凋亡诱导配体(TRAIL)蛋白对SKOV3移植瘤细胞半胱天冬氨酸蛋白酶-3(Caspase-3)表达的影响及其与肿瘤细胞凋亡的关系.方法 建立雌性裸小鼠SKOV3移植瘤24只,随机分为4组,每组6只.TRAIL组单用重组人TRAIL蛋白(10μg/kg),顺铂(DDP)组单用DDP(3 mg/...