乳腺癌新辅助化疗后细胞增殖与凋亡的变化及临床意义

目的：观察乳腺癌患者新辅助化疗前后细胞增殖及凋亡的变化，分析与化疗疗效的关系。方法：51例乳腺癌患者术前经Mammotome穿刺活检确诊，在CEF方案新辅助化疗前及化疗2个周期后分别进行癌组织中Ki-67的免疫组织化学染色和TUNEL原位细胞凋亡检测，并评价化疗疗效。结果：新辅助化疗前Ki-67阳性表达率及凋亡指数（AI）分别为23．2％和34．1％，化疗2个周期后Ki-67阳性表达率及A1分别为7．8％和67．5%，两者差异有统计学意义，P=0．004，P=0．006；化疗疗效临床CR5例，PR40例，SD6例，无疾病进展期病例，新辅助化疗疗效与Ki-67表达下降及肿瘤细胞凋亡增加显著相关。结论：新辅助化疗可明显抑制乳腺癌细胞的增殖，诱导凋亡，与化疗疗效密切相关。     

新辅助化疗诱导乳腺癌细胞凋亡与化疗疗效关系

[目的]探讨乳腺癌采用CEF方案新辅助化疗肿瘤组织标本中凋亡指数(apopto-sis index,AI)及其与化疗疗效的关系.[方法]确诊乳腺癌58例患者经予3个周期CEF方案新辅助化疗.化疗前后均检测AI值资料完整的42例.凋亡指数测定采用原位细胞末端转移标记法(TUNEL法).[结果]CEF方案化疗有效率为62.1%.化疗后AI值显著降低(P＜0.05).AI变化和术前化疗疗效密切相关(P＜0.05).[结论]化疗后凋亡指数高低可以作为预测新辅助化疗效果的指标.     

新辅助化疗对乳腺癌细胞凋亡和增殖的影响

背景与目的：在实验研究中,已证实多种化疗药物都可引起乳腺癌细胞的凋亡。但在人体内进行有关化疗药物引起乳腺癌细胞凋亡的研究,还少有报道。本文研究新辅助化疗能否引起乳腺癌肿瘤细胞的凋亡以及对乳腺癌肿瘤细胞增殖的影响。方法：应用末端转移酶介导的dUTP切口末端标记法（TdT-mediated dUTP nick end labelling,TUNEL)及免疫组化的标记链菌亲和素生物素法（Labelled Streptavidin Biotin,LSAB),分别检测100例乳腺癌组织中肿瘤细胞的凋亡指数（Apoptotic index,AI)和乳腺癌肿瘤组织的增殖细胞核抗原（Proliferating cell nuclear antigen,PCNA)表达。结果：新辅助化疗组肿瘤细胞凋亡指数（AI）均数为7．47％,与对照组肿瘤细胞凋亡指数（AI）均数4．83％相比明显增高（P＜0．01）。新辅助化疗组肿瘤细胞增殖细胞核抗原（proliferation cell nuclear antigen,PCNA)阳性表达率均数为33．71％,与对照组PCNA阳性率均数51．52％相比明显降低（P＜0．01）。在新辅助化疗组及对照组两组病例中,肿瘤细胞凋亡指数（AI）与增殖细胞核抗原（PCNA）的阳性表达均呈负相关。结论：在人体乳腺癌组织中,新辅助化疗能诱导肿瘤细胞发生凋亡,并能抑制其增殖。     

新辅助化疗对乳腺癌细胞凋亡和增殖的影响

目的：观察乳腺癌新辅助化疗后癌细胞凋亡及增殖程度的变化,判断化疗疗效.方法：新辅助化疗手术标本44例和术前未接受化疗的对照组37例,进行Ki67及PCNA免疫组织化学染色和TUNEL原位细胞凋亡检测,并进行统计学处理.结果：化疗组Ki67、PCNA和AI的阳性细胞数比率分别为11.8%、31.3%和69.8%,对照组分别为21.6%、62.8%和52.8%,化疗组和对照组相比均有显著性差异.结论：新辅助化疗能抑制乳腺癌细胞的增殖,诱导凋亡.     

新辅助化疗对乳腺癌肿瘤细胞的影响及其临床意义

目的:探索新辅助化疗对人体乳腺癌的作用机制及其临床应用的意义。方法:选取同期经病理确诊可手术的女性乳腺癌100例,分成新辅助化疗组和对照组各50例进行配对研究。新辅助化疗组术前接受CMF或CAF方案化疗2个疗程,对照组术前不接受任何抗肿瘤治疗。手术标本常规病理检查,同时应用原位末端标记法(TUNEL)及免疫组化LSAB法,分别检测乳腺癌组织中肿瘤细胞的凋亡指数(AI)和肿瘤组织的增殖细胞核抗原(PCNA)表达。结果:新辅助化疗组CR4例(8.0%),PR28例(56.0%),SD18例(36.0%),无恶化病例,总有效率为64.0%。新辅助化疗组AI(7.5%±2.9%),PCNA表达的阳性率(33.7%±6.8%);对照组AI(4.8%±2.1%),PCNA表达的阳性率(51.5%±10.2%)。AI与PCNA表达的阳性率均呈负相关,与化疗的疗效呈正相关。结论:乳腺癌新辅助化疗的总有效率(CR+PR)为64.0%;在人体乳腺癌组织中,新辅助化疗的作用机制除化疗药物的细胞毒作用外,还有可能通过诱导肿瘤细胞凋亡并抑制其增殖发挥抗肿瘤作用。     

新辅助化疗后乳腺癌组织中survivin、Ki-67和 AI的表达及其临床意义

目的探讨新辅助化疗后乳腺癌组织中survivin、Ki-67和AI的表达及其临床意义。方法采用免疫组化SABC法,检测60例行新辅助化疗和60例未行新辅助化疗的乳腺癌组织中survivin和Ki-67的表达,采用原位细胞末端转移标记法(TUNEL法)检测细胞凋亡。结果新辅助化疗组survivin和Ki-67阳性率分别为36.7%和38.3%,明显低于对照组(71.7%,61.7%),P均<0.05;新辅助化疗组AI均数为(9.34±3.12)%,低于对照组(5.27±3.16)%,P<0.05;新辅助化疗组survivin的表达与AI呈负相关,而与Ki-67表达呈正相关。新辅助化疗组化疗总有效率为73.3%,化疗后部分缓解者(Ⅱ级)病例survivin阳性率(18.2%)明显低于无效者(Ⅲ级)(81.3%),P<0.01。结论CTF方案新辅助化疗,可能通过抑制乳腺癌survivin表达而抑制肿瘤的增殖和诱导其凋亡。     

18F-FDG PET/CT融合显像对乳腺癌新辅助化疗疗效的预测价值

背景与目的:既往研究表明,18F-FDG PET显像结果与肿瘤新辅助治疗后的临床或病理反应密切相关.本研究拟探讨乳腺癌新辅助化疗前后18F-FDGPET/CT融合显像与细胞凋亡间的关系,以求进一步明确PET/CT对乳腺癌新辅助化疗疗效的预测价值.方法:45例原发性乳腺癌患者细针穿刺确诊后给予新辅助化疗三个周期,新辅助化疗前后行PET/CT融合显像检查并计算肿瘤区与非肿瘤区放射性比值(tumor to non-tumor activity ratio,T/N),dTUP末端标记技术检测穿刺及手术标本的癌细胞凋亡指数(apoptotic index,AI).结果:新辅助化疗后临床疗效评价完全缓解4例(8.9%),部分缓解29例(64.4%),疾病稳定10例(22.2%),疾病进展2例(4.4%).化疗前后肿瘤平均T/N值分别为3.23±0.63、2.31±0.49(P=0.006),下降6.4%～50.8%.化疗前后AI分别为(2.81±0.76)%、(17.31±6.85)%(P＜0.001),增高1.9%～41.3%.T/N值下降率与AI变化值间存在直线相关关系(r(.)=0.850,P＜0.001).以化疗后T/N值下降≥20%为阈值,PET/CT预测肿瘤临床缓解的灵敏度、特异度分别为90.9%、83.3%,阳性、阴性预测值分别为93.8%、76.9%,准确率为92.1%.结论:18F-FDG PET/CT融合显像与乳腺癌新辅助化疗后的细胞凋亡状态密切相关,对预测化疗疗效具有一定的应用价值.     

新辅助化疗对Ⅲ期非小细胞肺癌细胞凋亡和增殖的影响

目的：探讨新辅助化疗对非小细胞肺癌肿瘤细胞凋亡及对肿瘤细胞增殖的影响。方法：选取Ⅲ期非小细胞肺癌患者56例行新辅助化疗后并手术．并选取同期50例直接手术患者作为对照组。两组患者的手术标本．分别采用末端转移酶介导的dUTP切口末端标记法（TUNEL）行细胞凋亡检测，采用免疫组化标记链菌亲和素生物素法（LSAB），检测细胞增殖抗原Ki－67的表达。结果：新辅助化疗组肿瘤细胞凋亡指数（AI）均数为9．34％，对照组肿瘤细胞凋亡指数均数为5．27％．两组比较有显著差异（P〈0，001）。新辅助化疗组肿瘤细胞增殖抗原Ki－67阳性表达率均数为35．68％．对照组Ki－67阳性表达率均数为59.35％，两组比较差异显著（P〈0．001）。新辅助化疗组及对照组中，肿瘤细胞凋亡指数AI与增殖指数Ki-67的阳性表达均成负相关。结论：新辅助化疗能诱导Ⅲ期非小细胞肺癌肿瘤细胞的凋亡．并能抑制其增殖。     

细胞凋亡水平和宫颈癌化学治疗的研究回顾

通过对细胞凋亡水平和宫颈癌化学治疗的文献综述 ,阐明细胞凋亡指数 (apoptoticindex ,AI)和宫颈癌化疗的目前研究概况 ,以及细胞凋亡指数在宫颈癌化疗中的应用价值。目前结果表明 ,细胞凋亡指数在宫颈癌化疗 ,尤其是新辅助化疗中 ,具有广阔的临床运用前景。诱导细胞凋亡可能是宫颈癌化学治疗的机制之一。     

细胞凋亡水平和宫颈癌化学治疗的研究回顾

通过对细胞凋亡水平和宫颈癌化学治疗的文献综述,阐明细胞凋亡指数（apoptotic index,AI)和宫颈癌化疗的目前研究概况,以及细胞凋亡指数在宫颈癌化疗中的应用价值。目前结果表明,细胞凋亡指数在宫颈癌化疗,尤其是新辅助化疗中,具有广阔的临床运用前景。诱导细胞凋亡可能是宫颈癌化学治疗的机制之一。     

Prognostic role of p27Kip1 and apoptosis in human breast cancer

Human breast carcinoma is biologically heterogeneous, and its clinical course may vary from an indolent slowly progressive one to a course associated with rapid progression and metastatic spread. It is important to establish prognostic factors which will define subgroups of patients with low vs high risk of recurrence so as to better define the need for additional therapy. Additional characterization of the molecular make-up of breast cancer phenotypes should provide important insights into the biology of breast cancer. In the present study, we investigated apoptosis, expression of p27Kip1 and p53 retrospectively in 181 human breast cancer specimens. In addition, their relevance to the biological behaviour of breast cancer was examined. Our studies found a significant association among high histological grade, high p53, low apoptosis and low p27. Our results also demonstrated that, in human breast cancer, low levels of p27 and apoptotic index (AI) strongly correlated with the presence of lymph node metastasis and decreased patient survival. In node-negative patients, however, p27 also had prognostic value for relapse-free and overall survival in multivariate analysis. Furthermore p27 and AI had predictive value for the benefits of chemotherapy. These latter observations should prompt prospective randomized studies designed to investigate the predictive role of p27 and AI in determining who should receive chemotherapy in node-negative patients.     

Reduced apoptosis and proliferation and increased Bcl-2 in residual breast cancer following preoperative chemotherapy

Experimental laboratory data suggest that tumour growth is a balance between apoptosis and proliferation and that suppression of drug-induced apoptosis by oncogenes such as bcl-2 may be an important cause of intrinsic chemoresistance. The aims of this study were to assess the in vivo relationship of apoptosis to proliferation and Bcl-2 protein in human breast tumours both prior to chemotherapy and in the residual resistant cell population at the completion of treatment. We examined apoptotic index (AI), Ki67 and Bcl-2 protein expression in the tissue of 40 patients with operable breast cancer immediately before ECF preoperative chemotherapy, and in 20 of these patients with residual tumour, at the completion of treatment. There was a significant positive association between AI and Ki67 both before and after chemotherapy, and in their percentage change with treatment. In the residual specimens AI and Ki67 were significantly reduced compared with pre-treatment biopsies, while Bcl-2 expression showed a significant increase. No differences were seen in the pre-treatment levels of any of the variables measured between patients obtaining pathological complete response and those who did not, although numbers were small. These data suggest that apoptosis and proliferation are closely related in vivo. It is possible that the phenotype of reduced apoptosis and proliferation, and increased Bcl-2 may be associated with breast cancer cells resistant to cytotoxic chemotherapy, although this can only be proven by assessing larger numbers of patients in relation to pathological response.     

Co-transduction of Apaf-1 and caspase-9 augments etoposide-induced apoptosis in U-373MG glioma cells.

Several apoptosis-related genes have been reported to be involved in chemotherapy-induced apoptosis in cancers. An assessment of the relationship between expression of those genes and the degree of chemotherapy-induced apoptosis may be useful in improving the efficacy of cancer therapy. We transduced Apaf-1 (apoptotic protease-activating factor-1) and caspase-9 into U-373MG glioma cells using adenovirus (Adv) vectors in the presence of etoposide and evaluated the degree of apoptosis. The degree of apoptosis in etoposide-treated U-373MG cells infected with Adv for Apaf-1 (Adv-APAF1) was higher (27%) than that in cells infected with control Adv (14%), that in cells infected with Adv for caspase-9 (Adv-Casp9) was higher (34%) than that in cells infected with Adv-APAF1, and that in cells infected with both Adv-APAF1 and Adv-Casp9 was the highest (41%). Treatment with etoposide increased expression of p53 and decreased expression of Bcl-X(L) in U-373MG cells which harbored mutant p53. These results indicate that the expression of Apaf-1 and caspase-9 may be important determinants in predicting the sensitivity of cancers to chemotherapy. Adv-mediated co-transduction of Apaf-1 and caspase-9 should render cancer cells highly sensitive to chemotherapy.     

Co-transduction of Apaf-1 and Caspase-9 Augments Etoposide-induced Apoptosis in U-373MG Glioma Cells

Several apoptosis-related genes have been reported to be involved in chemotherapy-induced apoptosis in cancers. An assessment of the relationship between expression of those genes and the degree of chemotherapy-induced apoptosis may be useful in improving the efficacy of cancer therapy. We transduced Apaf-1 (apoptotic protease-activating factor-1) and caspase-9 into U-373MG glioma cells using adenovirus (Adv) vectors in the presence of etoposide and evaluated the degree of apoptosis. The degree of apoptosis in etoposide-treated U-373MG cells infected with Adv for Apaf-1 (Adv-APAFl) was higher (27%) than that in cells infected with control Adv (14%), that in cells infected with Adv for caspase-9 (Adv-Casp9) was higher (34%) than that in cells infected with Adv-APAFl, and that in cells infected with both Adv-APAFl and Adv-Casp9 was the highest (41%). Treatment with etoposide increased expression of p53 and decreased expression of Bcl-X_L in U-373MG cells which harbored mutant p53. These results indicate that the expression of Apaf-1 and caspase-9 may be important determinants in predicting the sensitivity of cancers to chemotherapy. Adv-mediated co-transduction of Apaf-1 and caspase-9 should render cancer cells highly sensitive to chemotherapy.     

Reduced Apaf-1 expression in human cutaneous melanomas

Malignant melanoma is a life-threatening skin cancer due to its highly metastatic character and resistance to radio- and chemotherapy. It is believed that the ability to evade apoptosis is the key mechanism for the rapid growth of cancer cells. However, the exact mechanism for failure in the apoptotic pathway in melanoma cells is unclear. p53, the most frequently mutated tumour suppressor gene in human cancers, is a key apoptosis inducer. However, p53 mutation is only found in 15-20% of melanoma biopsies. Recently, it was found that Apaf-1, a downstream target of p53, is inactivated in metastatic melanoma. Specifically, loss of heterozygosity (LOH) of the Apaf-1 gene was found in 40% of metastatic melanoma. To determine if loss of Apaf-1 expression is indeed involved in melanoma progression, we employed the tissue microarray technology and examined Apaf-1 expression in 70 human primary malignant melanoma biopsies by immunohistochemistry. Our data showed that Apaf-1 expression is significantly reduced in melanoma cells compared with normal nevi (chi(2)=6.02, P=0.014). Our results also revealed that loss of Apaf-1 was not associated with the tumour thickness, ulceration or subtype, patient's gender, age and 5-year survival. In addition, our in vitro apoptosis assay revealed that overexpression of Apaf-1 can sensitise melanoma cells to anticancer drug treatment. Taken together, our data indicate that Apaf-1 expression is significantly reduced in human melanoma and that Apaf-1 may serve as a therapeutic target in melanoma.     

Over-expression of APAF-1 and caspase-9 augments radiation-induced apoptosis in U-373MG glioma cells.

The p53 tumor-suppressor gene plays a critical role in radiation-induced apoptosis. Several genes, including Bax and Fas, are involved in p53-mediated apoptosis, and their over-expression enhances the degree of radiation-induced apoptosis. Apaf-1 and caspase-9 have been reported to be downstream components of p53-mediated apoptosis, suggesting that these genes play a role in radiation-induced apoptosis. In this study, we transduced U-373MG cells harboring mutant p53 with the Apaf-1 and/or caspase-9 genes via adenoviral (Adv) vectors concomitant with X-ray irradiation and evaluated the degree of apoptosis. The percentage of apoptotic cells in U-373MG cells co-infected with the Adv for Apaf-1 (Adv-APAF-1) and that for caspase-9 (Adv-Casp9) and treated with irradiation (24%) was much higher than that in cells co-infected with Adv-APAF-1 and Adv-Casp9 and not treated with irradiation (0.86%) and that in cells infected with either Adv-APAF-1 or Adv-Casp9 and treated with irradiation (2.0% or 2.6%, respectively). The apoptosis induced by co-transduction of Apaf-1 and caspase-9 and irradiation was repressed in cells that were co-infected with the Adv for Bcl-X(L) but not in cells co-infected with the Adv for Bcl-2. These results indicate that Apaf-1 and caspase-9 play a role in radiation-induced apoptosis in cancer cells harboring mutant p53. Bcl-X(L) may be critically involved in the radioresistance of cancer cells by repressing Apaf-1- and caspase-9-mediated apoptosis. Expression of Apaf-1 and caspase-9 in tumors may be an important determinant of the therapeutic effect of irradiation in cancer treatment.     

Inhibition of the proliferation of acquired aromatase inhibitor-resistant breast cancer cells by histone deacetylase inhibitor LBH589 (panobinostat)

Aromatase inhibitors (AIs) are important drugs for treating postmenopausal patients with hormone receptor-positive breast cancer. However, acquired resistance to AI therapies is a significant problem. Our study has revealed that the histone deacetylase inhibitor LBH589 treatment abrogated growth of AI-resistant cells in vitro and in vivo, causing cell cycle G2/M arrest and induced apoptosis. LBH589 treatment also reduced the level of NF-κB1 which is overexpressed when AI resistance develops. Analyzing paired tumor specimens from 12 patients, we found that NF-κB1 expression was increased in recurrent AI-resistant tumors as compared to the paired primary tumors before AI treatment. This finding was consistent with up-regulated NF-κB1 expression seen in a collection of well-established AI-resistant cell lines. Furthermore, knockdown of NF-κB1 expression significantly suppressed the proliferation of AI-resistant cells. Treatment of AI-resistant cell lines with LBH589 suppressed NF-κB1 mRNA and protein expression. In addition, LBH589 treatment abrogated growth of AI-resistant tumors in mice, and was associated with significantly decreased levels of NF-κB1 in tumors. In all, our findings strongly support further investigation of LBH589 as a novel therapeutic strategy for patients with AI-resistant breast cancer, in part by suppressing the NF-κB1 pathway.     

Co-transduction of Apaf-1 and caspase-9 highly enhances p53-mediated apoptosis in gliomas

Mutation of the p53 gene plays a critical role in the development of cancer and response to cancer therapy. To analyze the mechanism of cancer development and to improve cancer therapy, it is important to assess which genes are downstream components of p53 in cancers, and whether the expression levels of these genes affect p53-mediated apoptosis. In this study, we transduced the wild type p53 gene along with the Apaf-1 and caspase-9 genes via adenovirus vectors into U251 and U-373MG glioma cells harbouring a mutated p53, and evaluated the degree of apoptosis. Co-induction of Apaf-1 and caspase-9 genes highly enhanced p53-mediated apoptosis in glioma cells. Induction of wild type p53 enhanced the expression levels of Bax, p21/WAF1, and Fas protein. To determine which gene is activated by wild type p53 induction and, in turn, activates Apaf-1 and caspase-9, we transduced the Bax, p21/WAF1 or Fas gene via adenovirus vector to U251 cells to achieve a similar expression level as that induced by the Adv for p53 in U251 cells. U251 cells transduced with Fas concomitant with the Apaf-1 and caspase-9 genes underwent drastic apoptosis. This suggests that induction of wild type p53 upregulates Fas, which in turn may play a role in the activation of Apaf-1 and caspase-9. These results are important for analyzing the mechanism of tumour development and for predicting the therapeutic effect of p53 replacement gene therapy in a particular patient.     

Neo-adjuvant chemotherapy for operable breast cancer induces apoptosis

The use of neo-adjuvant chemotherapy (often referred to as pre-operative or primary chemotherapy) represents a major change in the management of breast cancer as a systemic disease. Laboratory studies have shown that many anti-cancer agents with differing modes of action achieve cytotoxic effects by inducing apoptosis. In this study, we investigated the induction of apoptosis by neo-adjuvant chemotherapy in human breast cancer. The aim was to determine whether a correlation existed between post chemotherapy apoptotic index (AI) and clinical response and patients' survival. Our results indicate that apoptosis is induced by neo-adjuvant chemotherapy and that the response is variable. Our data show that post chemotherapy AI correlated with clinical response and increased patient survival, including both relapse (disease) free survival and overall survival. Post-neo-adjuvant chemotherapy AI levels in primary breast cancer may possibly predict an individual patient's overall response.