重组人p53腺病毒联合治疗晚期恶性肿瘤的临床观察

目的：探讨重组人p53腺病毒注射液（rAd-p53）治疗恶性肿瘤的疗效及安全性。方法：对67例晚期无法行手术切除的恶性肿瘤患者,或术后肿瘤转移、无法再手术的患者,采用瘤内注射、血管介入和静脉滴注rAd-p53联合化疗和放疗,至少1个疗程以上,通过临床观察、KPS评分、CT或MRI检查以及随访,评价rAd-p53联合化、放疗的临床疗效。结果：67例患者治疗有效率为47.8%,疾病控制率为89.6%,其中CR患者4例,分别为卵巢癌1例,皮肤癌1例,鼻咽癌2例。rAd-p53联合放疗41例,联合化疗26例,有效率分别为56.1%和34.6%（P〈0.05）,疾病控制率分别为95.1%和80.8%（P〈0.05）。全组中位生存时间为60个月（6～117个月）;联合治疗后,全组中位生存时间为24个月（2～34个月）。不良反应主要为自限性发热,另有个别胃肠道反应及肌肉酸痛。结论：rAd-p53对多种恶性肿瘤具有一定疗效,临床应用安全;局部给药联合放疗可能比联合化疗更能提高肿瘤的局控率。     

重组人p53腺病毒注射液局部治疗耐药舌癌1 例及分析

重组人p53腺病毒注射液是由5型腺病毒载体与人p53基因重组的肿瘤基因制品,我院局部治疗耐药舌癌1例,疗效显著.     

重组人 p53 腺病毒在晚期卵巢癌治疗中临床疗效及安全性评价简

目的：探讨重组人p53腺病毒注射液（rAd p53）联合TP／TC静脉化疗方案在晚期卵巢癌患者中的应用价值。方法：47例晚期卵巢癌术后患者,25例对照组患者单纯接受3周一次的TP／TC静脉化疗,共6—8疗程;22例实验组患者在化疗基础上加用2疗程rAd p53,比较2组患者近期、远期疗效及药物的安全性。结果：实验组CA125下降曲线显于对照组（P=0．037）;实验组无瘤生存期优于对照组（30．4个月vs21．5个月,P=0．012）;临床缓解率两组无差异（68．2％vs48．0％,P=0．135）;总生存期无差异（39．6个月vs32．5个月,P=0．13）。未发现rAd p53的严重不良反应。结论：rAd p53对于晚期卵巢癌患者的治疗有积极的作用,可以被很好的耐受,但结果尚需要进一步在临床应用中得以验证。     

重组人p53腺病毒注射液联合顺铂治疗肺癌所致胸腔积液的临床研究

背景与目的：肿瘤抑制基因p53是目前研究最为广泛和系统的抑癌基因之一,p53基因突变或缺失导致肿瘤的形成。本研究评价重组人p53腺病毒注射液（rAd-p53）导入野生型p53基因抑癌基因治疗同时联合顺铂治疗肺癌所致胸腔积液的临床疗效和毒副反应。方法：将35例肺癌合并胸腔积液患者随机分为联合组和单药组两组。所有患者应用长春瑞滨25mg/m2静脉点滴,第1、8天,每3周重复一次。前述治疗基础上,联合组胸腔内灌入rAd-p531×10^12VP和顺铂注射液40mg/m2;单药组胸腔内灌入顺铂注射液40mg/m2,每周重复一次,连用4次后观察疗效。结果：联合组和单药组的有效率分别为82.35%和50.00%（P〈0.05）;联合组和单药组的一般状况改善率分别为64.70%和33.33%（P〈0.05）;两组患者主要不良反应为发热、胸痛、消化道反应及白细胞减少,联合组发热的发生率高于单药组（P〈0.05）,主要为自限性发热,36h后自行恢复正常。结论：重组人p53腺病毒注射液联合顺铂治疗肺癌所致胸腔积液疗效确切,且毒副反应较低,值得临床推广应用。     

重组人p53腺病毒联合化疗对复发性卵巢癌临床疗效的评价

目的：探讨重组人p53腺病毒注射液（rAd-p53）联合静脉化疗对复发性上皮性卵巢癌（recurrent epithelial ovarian carcinoma,REOC）的临床疗效。方法：选取甘肃省妇幼保健院妇产科2007年7月至2011年12月住院的49例REOC患者,其中25例在静脉化疗基础上加用rAd-p53治疗（联合组）,24例单纯进行静脉化疗（化疗组）,比较两组患者近期、远期疗效及药物的不良反应。结果：3个疗程治疗后,联合组CA125下降正常率高于化疗组（68.0% vs 33.3%,P=0.02）;疾病控制率两组分别为92.0%与87.5%,差异无统计学意义（χ2=1.03,P=0.59）。随访5~41个月（中位时间26个月）,两组患者总生存期无明显差异（18.8个月vs 13.9个月,P=0.051）。患者使用rAd-p53未发现严重的不良反应。结论：rAd-p53联合化疗对于复发性REOC患者似乎有一定受益,无明显的不良反应,结果尚需要进一步在临床应用中得以验证。     

重组人p53基因腺病毒注射液在原发性肝癌介入治疗中的临床研究

目的:研究重组人p53腺病毒注射液（商品名:今又生）在原发性肝癌介入治疗中的安全性及有效性。方法:61例中晚期原发性肝癌患者,分两组介入治疗:对照组:碘油+化疗药（5-FU+EPI+DDP）;治疗组:碘油+化疗药（5-FU+EPI+DDP）+重组人p53基因腺病毒注射液;每4周一次,连用3次。结果:不论是对照组还是治疗组,毒副反应不大,耐受性良好,生活质量均有一定的提高;治疗后两组均能使原发性肝癌的肿瘤标志物AFP下降,但治疗组下降明显;治疗组的有效率（82.76%）明显高于对照组（59.38%）,统计学检验差异显著。结论:今又生介入治疗原发性肝癌安全有效,值得推广。     

重组人p53腺病毒治疗原发性肝癌的血清p53蛋白检测及其临床意义

目的:初步观察重组人p53腺病毒注射液(recombinant adenovirus-p53,rAd-p53)治疗肝癌临床疗效,和治疗前后血清p53蛋白的变化及其临床意义。方法:对38例原发性肝癌患者,采用瘤内注射、血管介入和静脉滴注rAd-p53联合化疗和放疗,行至少1个疗程以上的治疗,治疗结束一个月后评价疗效。应用ELISA检测患者治疗前和治疗后血清p53蛋白。结果:38例患者总的疾病控制率为94.74%,其中CR7例,PR11例,SD18例,PD2例。38例肝癌患者经人p53腺病毒注射液治疗后血清p53蛋白含量比治疗前明显升高(P<0.01)。治疗前血清p53蛋白较高的患者比p53蛋白低的对人p53腺病毒注射液的治疗反应较好。结论:人p53腺病毒注射液治疗对肝癌具有一定疗效,临床应用安全;治疗前血清p53蛋白含量以及动态变化可能是预测rAd-p53治疗肝癌疗效的有价值指标,并对人p53腺病毒注射液的临床应用具有一定的指导作用。     

重组人p53 腺病毒注射液联合新辅助化疗治疗局部晚期子宫颈癌的临床观察

 目的　评价重组人p53腺病毒（rhAd-p53）注射液联合新辅助化疗治疗局部晚期子宫颈癌的疗效及安全性。方法　以收治的ⅠB2～ⅢA期局部晚期子宫颈癌患者40例为研究对象,采用简单随机化分组方法分为两组：单纯静脉PVB方案化疗组（PVB组）20例,rhAd基因治疗+静脉PVB方案化疗组（rhAd-p53+PVB组）20例。PVB组PVB方案化疗1个疗程;rhAd-p53+PVB组化疗方案相同,于化疗后第3天第1次注射rhAd-p53制剂1×1012 VP病毒颗粒,然后每3 d 1次,共3次。观察肿瘤体积的变化、患者的不良反应和肿瘤组织中血管内皮生长因子（VEGF）、p53蛋白和微血管密度（MVD）水平,对照组为未用放化疗及基因治疗的子宫颈癌患者。结果　所有患者化疗结束3周后评价疗效,PVB组用药后肿瘤缩小（11.42±2.78）cm2,rhAd-p53+PVB组肿瘤缩小（15.25±4.00）cm2,差异有统计学意义（P＜0.05）。PVB组有效率为75 %（15／20）,rhAd-p53+ PVB组有效率为95 %（19／20）。VEGF、p53蛋白、MVD在对照组、PVB组、rhAd-p53+PVB组中的阳性表达率均呈逐渐降低的趋势,差异有统计学意义（P＜0.05）。基因治疗联合化疗未增加化疗患者的不良反应。结论　子宫颈癌患者瘤内注射rAd-p53制剂是有效的,rhAd-p53制剂是子宫颈癌的一种有潜力的基因治疗药物。     

Tf-PEG脂质体-rAdp53复合物、rAdp53腹腔内灌注治疗晚期结直肠癌合并恶性腹腔积液的临床研究

目的:观察Tf-PEG脂质体-rAdp53复合物、rAdp53腹腔内灌注治疗晚期结直肠癌合并恶性腹腔积液临床疗效。方法:采用Tf-PEG脂质体-rAdp53复合物、rAdp53治疗晚期结直肠癌合并恶性腹腔积液患者,随机分为3组,每组22例。观察组:制备Tf-PEG脂质体-rAdp53复合物,溶于20ml生理盐水后注入腹腔,再缓慢灌入40℃的灭菌用水1000ml+地塞米松10mg。对照组:将1支rAdp53(1×1012VP)+灭菌用水1000ml+地塞米松10mg混合后腹腔灌注。空白组:灭菌用水1000ml+地塞米松10mg腹腔灌注。4周后评价其疗效及不良反应。结果:观察组总有效率90.90%(20/22);对照组总有效率72.73%(16/22);空白组总有效率22.72%(5/22)。观察组和对照组有效率显著高于空白组,且观察组有效率亦显著高于对照组(P<0.05)。观察组和对照组中获RR患者Karnofsky评分较治疗前明显提高且腹围减小。结论:Tf-PEG脂质体-rAdp53复合物较rAdp53在腹腔灌注治疗晚期结直肠癌合并恶性腹腔积液的疗效更明显,不良反应较小。     

手术联合重组人p53腺病毒注射液治疗复发性恶性胶质瘤

目的 探讨手术联合重组人p53腺病毒注射液(rAd-p53)治疗复发性恶性胶质瘤的安全性和有效性.方法 38例复发性恶性胶质瘤患者再次行手术治疗.联合治疗组的18例患者术中埋放Ommaya储液囊,将1×1012VP rAd-p53经Ommaya储液囊注入瘤腔,每周1次,4周为1个疗程,连续2～4个疗程.手术组20例患者单纯行手术切除.两组患者均随访1年以上.结果 联合治疗组6个月生存率为66.7%,1年生存率为44.4%,中位生存期为9.7个月,较手术组(7.1个月)明显提高.联合治疗组术后6个月Karnofsky评分为(81.8±15.7)分,较术前明显提高(P＜0.05);术后1年Karnofsky评分为(66.1±17.4)分,明显高于手术组[(53.5±18.2)分,P＜0.05].结论 rAd-p53对于复发性恶性胶质瘤安全有效,能延长患者生存期,改善患者的生活质量.     

Clinical study of adenoviral mediated p53 gene therapy for non-small cell lung cancer in Japan

The prognosis in case of non-small cell lung cancer (NSCLC) remains poor, and novel treatment modalities are urgently needed for advanced NSCLC. Backed by advances in the understanding of cancer biology, gene therapy has been developed in recent years. The p53 gene is altered in over 50% of cancers and has been extensively studied as a tumor suppressor gene. Adenoviral-mediated p53 gene transfer is currently under clinical evaluation worldwide for the treatment of cancer. We are now conducting a phase I study of Ad-p53 for advanced NSCLC patients in Japan. As an interim report, we provide a brief summary of the current status of this study, highlighting the safety and clinical efficacy of Ad-p53. As of September 2002, 13 patients were enrolled to this study, and safety and antitumor effects have been noted.     

Adenoviral p53 gene therapy for human lung cancer

Recent advances in molecular biology have fostered remarkable insights into the molecular basis of neoplasms. This new understanding of cancer pathogenesis suggests that restoration of the function of critical gene products could halt or reverse these mechanisms, thus having a therapeutic effect in cancer. The tumor suppressor p53 gene has been implicated in many inherited and sporadic forms of malignancy in humans. A number of preclinical experiments have demonstrated that restoration of the wild-type p53 function in the cancer cell by gene transfer is sufficient to cause antitumor effects such as cell-cycle arrest and induction of apoptosis. This approach has entered initial clinical testing and provided intriguing information about the intratumoral administration of an adenovirus vector expressing the wild-type p53 gene in non-small cell lung cancer patients.     

Cooperative effect of adenoviral p53 gene therapy and standard chemotherapy in ovarian cancer cells independent of the endogenous p53 status

Clinical adenoviral p53 gene therapy has been shown by us and others to inhibit tumor growth of ovarian cancer with endogenous mutant p53. This study was designed to test the cooperative antitumor effect of standard combination chemotherapy using paclitaxel and carboplatin together with adenoviral p53 gene transfer in the presence of wild-type and mutant p53. Seven ovarian cancer cell lines with mutant p53 and seven ovarian cancer cell lines with wild-type p53 were tested. An E1-deleted adenovirus type 5 expressing p53 (ACNp53) was used for p53 gene transfer. p53 gene transfer at 50% transduction efficiency significantly reduced IC50 of carboplatin chemotherapy up to 49-fold, of paclitaxel chemotherapy up to six-fold, and of paclitaxel/carboplatin chemotherapy up to 19-fold in the wild-type p53 cell line OV-MZ-5. Synergism between ACNp53 and chemotherapy calculated by median-effect analysis was found at low drug concentrations in all cell lines independent of the p53 mutational status. In conclusion, adenoviral p53 gene transfer significantly increased the sensitivity of ovarian tumor cells to paclitaxel, to carboplatin and/or to the combination of both.     

Phase I/II adenoviral p53 gene therapy for chemoradiation resistant advanced esophageal squamous cell carcinoma

We investigated the feasibility, safety, biological activity and therapeutic efficacy of adenovirus-mediated p53 gene transfer in patients with chemoradiation resistant advanced esophageal carcinoma. Eligible patients were not surgical candidates and had measurable, advanced squamous cell carcinoma of the esophagus that was resistant to chemoradiation therapy. On a 28-day cycle, intratumoral injections of Ad5CMV-p53 (INGN 201; ADVEXIN) were administered on days 1 and 3 at four dose levels (10 x 10(11) particles to 25 x 10(11) particles) and treated for up to five cycles. Ten patients received a total of 26 cycles with no dose-limiting toxicity. Administration of multiple courses was feasible and well-tolerated. Local tumor responses revealed stable disease in nine cases and progressive disease in one case. The overall responses were stable in six and progressive in four cases. Using polymerase chain reaction (PCR) analyses, gene transfer and p53 specific transgene expression were detected in tumor biopsy tissue from all patients. mRNA levels of p53, p21 and MDM2 increased in all but one case. Three patients showed absence of disease upon repeat biopsies. Substantial improvement in swallowing was observed in one patient with stenotic lesions. Intratumoral injection of Ad5CMV-p53 is safe, feasible and biologically active when administered in multiple doses to patients with esophageal cancer. Observations from this study indicate that this treatment results in local antitumor effects in chemoradiation resistant esophageal squamous cell carcinoma.     

腺病毒介导的p53基因对喉癌细胞生长的抑制作用

目的探索ｐ５３基因在喉癌基因治疗方面的可行性。方法以人喉癌细胞系Ｈｅｐ－２为实验对象,将载有人野生型ｐ５３ｃＤＮＡ并含巨细胞病毒（ＣＭＶ）启动子的重组腺病毒（Ａｄ５ＣＭＶ－ｐ５３）感染Ｈｅｐ－２细胞及肿瘤组织,体内外实验观察Ａｄ５ＣＭＶ－ｐ５３对Ｈｅｐ－２细胞生长的影响。结果当Ａｄ５ＣＭＶ－ｐ５３在１００ＭＯＩ效靶比时,全部Ｈｅｐ－２细胞得到转染。感染２天后ｐ５３蛋白表达达到高峰,Ｈｅｐ－２生长受到明显的抑制。Ａｄ５ＣＭＶ－ｐ５３感染Ｈｅｐ－２细胞在裸鼠中失去致瘤性。瘤内注射Ａｄ５ＣＭＶ－ｐ５３后,荷瘤裸鼠的肿瘤体积明显减小。结论Ａｄ５ＣＭＶ－ｐ５３转导野生型ｐ５３基因可能是一种有效的喉癌基因治疗途径。     

Viral shedding after p53 adenoviral gene therapy in 10 cases of esophageal cancer

We detected adenoviral DNA fragments in excretions of 10 esophageal cancer patients by DNA-PCR after tumor injection of Ad-CMV-vector. A total of 220 samples consisting of feces, gargling saliva, urine, and blood plasma were assessed. A total of 29.7% of feces samples and 13.2% of gargling saliva samples were positive for adenoviral DNA fragments, but 89.7% of the positive feces samples and all of the positive gargling saliva samples turned negative on day 12 after tumor injection. Although adenoviral DNA fragments may be pathogen-free, patients' feces and gargling saliva contain adenoviral DNA fragments for 12 days after injection. ( Cancer Sci 2010; 101: 289–291)     

The evaluation of adenoviral p53-mediated bystander effect in gene therapy of cancer.

Because many tumors have mutated p53, one potential strategy proposed for cancer gene therapy is the introduction of the wild-type p53 gene into tumor cells. One puzzling aspect of this approach is that currently available gene transfer protocols result in a small percentage of tumor cells being transduced in vivo, thus implicating a "bystander effect" to achieve therapeutic efficacy. Because bystander effects in the context of p53-mediated gene therapy have not been well characterized, we evaluated the role of in vitro and in vivo bystander effects of adenovirally delivered p53 (AdWTp53). Using human tumor cell lines that did not express p53 protein but were infectible with adenovirus and showed sensitivity to p53-mediated apoptosis, we were unable to demonstrate an AdWTp53-mediated in vitro bystander effect, despite seeing strong bystander effects when cells were infected with an adenovirus containing the suicide gene herpes simplex virus thymidine kinase and treated with ganciclovir. In contrast, in vivo flank mixing studies using one of these cell lines showed a weak but significant p53-mediated bystander effect (a 40% inhibition of tumor growth). This bystander effect translated into a small survival advantage in an established intraperitoneal tumor model when tumor burden was low at the time of viral instillation. The survival advantage was lost, however, when tumor burden was increased. This study indicates that treatment of human tumors using AdWTp53 may be possible; however, because of the weak bystander effect in vivo, effective treatment will likely require a large percentage of tumor cells to be transduced.     

Intraperitoneal gene therapy with adenoviral-mediated p53 tumor suppressor gene for ovarian cancer model in nude mouse

In an effort to develop a method for better local control of advanced ovarian cancers, we have established a peritoneal tumor model of ovarian cancer in the nude mouse and applied intraperitoneal gene therapy with the recombinant adenoviral-mediated wild-type p53 tumor suppressor gene (Avp53). The results indicate that: (a) the recombinant adenoviral vector system effectively infected the tumor and normal cells in the peritoneal cavity; and (b) Avp53 treatment effectively suppressed the growth of peritoneal tumors and prolonged the survival of the treated group, especially when the tumor burden was less. These results suggest that intraperitoneal gene therapy using Avp53 is potentially useful as an adjuvant therapeutic modality in human ovarian cancer.