Department of Gynecologic Oncology, University of Texas M. D. Anderson Cancer Center, Houston, TX 77230-1439, USA.
J Natl Cancer Inst. 2011 Nov 2;103(21):1596-612. doi: 10.1093/jnci/djr372. Epub 2011 Sep 28.
We investigated the clinical and biological significance of p130cas, an important cell signaling molecule, in ovarian carcinoma.
Expression of p130cas in ovarian tumors, as assessed by immunohistochemistry, was associated with tumor characteristics and patient survival. The effects of p130cas gene silencing with small interfering RNAs incorporated into neutral nanoliposomes (siRNA-DOPC), alone and in combination with docetaxel, on in vivo tumor growth and on tumor cell proliferation (proliferating cell nuclear antigen) and apoptosis (terminal deoxynucleotidyl transferase dUTP nick-end labeling) were examined in mice bearing orthotopic taxane-sensitive (HeyA8 and SKOV3ip1) or taxane-resistant (HeyA8-MDR) ovarian tumors (n = 10 per group). To determine the specific mechanisms by which p130cas gene silencing abrogates tumor growth, we measured cell viability (MTT assay), apoptosis (fluorescence-activated cell sorting), autophagy (immunoblotting, fluorescence, and transmission electron microscopy), and cell signaling (immunoblotting) in vitro. All statistical tests were two-sided.
Of 91 ovarian cancer specimens, 70 (76%) had high p130cas expression; and 21 (24%) had low p130cas expression. High p130cas expression was associated with advanced tumor stage (P < .001) and higher residual disease (>1 cm) following primary cytoreduction surgery (P = .007) and inversely associated with overall survival and progression-free survival (median overall survival: high p130cas expression vs low expression, 2.14 vs 9.1 years, difference = 6.96 years, 95% confidence interval = 1.69 to 9.48 years, P < .001; median progression-free survival: high p130cas expression vs low expression, 1.04 vs 2.13 years, difference = 1.09 years, 95% confidence interval = 0.47 to 2.60 years, P = .01). In mice bearing orthotopically implanted HeyA8 or SKOV3ip1 ovarian tumors, treatment with p130cas siRNA-DOPC in combination with docetaxel chemotherapy resulted in the greatest reduction in tumor growth compared with control siRNA therapy (92%-95% reduction in tumor growth; P < .001 for all). Compared with control siRNA therapy, p130cas siRNA-DOPC reduced SKOV3ip1 cell proliferation (31% reduction, P < .001) and increased apoptosis (143% increase, P < .001) in vivo. Increased tumor cell apoptosis may have persisted despite pan-caspase inhibition by the induction of autophagy and related signaling pathways.
Increased p130cas expression is associated with poor clinical outcome in human ovarian carcinoma, and p130cas gene silencing decreases tumor growth through stimulation of apoptotic and autophagic cell death.
我们研究了 p130cas 这种重要的细胞信号分子在卵巢癌中的临床和生物学意义。
通过免疫组织化学评估 p130cas 在卵巢肿瘤中的表达，将其与肿瘤特征和患者生存情况相关联。用小干扰 RNA（siRNA）掺入中性纳米脂质体（siRNA-DOPC）沉默 p130cas 基因，单独或与多西紫杉醇联合，对荷有原位紫杉醇敏感（HeyA8 和 SKOV3ip1）或紫杉醇耐药（HeyA8-MDR）卵巢肿瘤的小鼠（每组 n = 10）的体内肿瘤生长及肿瘤细胞增殖（增殖细胞核抗原）和凋亡（末端脱氧核苷酸转移酶 dUTP 缺口末端标记）进行检测。为了确定 p130cas 基因沉默阻断肿瘤生长的具体机制，我们在体外测量细胞活力（MTT 测定）、凋亡（荧光激活细胞分选）、自噬（免疫印迹、荧光和透射电子显微镜）和细胞信号（免疫印迹）。所有统计检验均为双侧检验。
在 91 例卵巢癌标本中，有 70 例（76%）有高 p130cas 表达；有 21 例（24%）有低 p130cas 表达。高 p130cas 表达与晚期肿瘤分期（P <.001）和初次细胞减灭术后残留病灶（>1 cm）（P =.007）有关，而与总生存期和无进展生存期呈负相关（中位总生存期：高 p130cas 表达 vs 低表达，2.14 对 9.1 年，差异为 6.96 年，95%置信区间为 1.69 至 9.48 年，P <.001；中位无进展生存期：高 p130cas 表达 vs 低表达，1.04 对 2.13 年，差异为 1.09 年，95%置信区间为 0.47 至 2.60 年，P =.01）。在荷有原位植入 HeyA8 或 SKOV3ip1 卵巢肿瘤的小鼠中，与对照 siRNA 治疗相比，用 p130cas siRNA-DOPC 联合多西紫杉醇化疗治疗导致肿瘤生长的减少最大（肿瘤生长减少 92%-95%；P <.001 所有）。与对照 siRNA 治疗相比，p130cas siRNA-DOPC 降低了 SKOV3ip1 细胞的增殖（减少 31%，P <.001）和增加了凋亡（增加 143%，P <.001）。肿瘤细胞凋亡的增加可能是由于诱导自噬和相关信号通路而导致全细胞凋亡抑制。
p130cas 在人类卵巢癌中的高表达与不良临床预后相关，p130cas 基因沉默通过刺激细胞凋亡和自噬性细胞死亡来降低肿瘤生长。
