康莱特注射液对肝癌化疗增敏作用的实验研究

目的：研究康莱特（KLT）注射液对肝癌细胞化疗增敏作用。方法：用癌细胞药物敏感性试验（MTT法）测定KLT注射液和5种化疗药联合作用的敏感度与对照组比较。结果：KLT注射液联合化疗药的抑瘤率高于单纯化疗药（P＜0．05）。结论：KLT注射液分别和5－FU、CP、DDP、MMC联用比单纯化学药物治疗肝癌有明显的增敏作用。     

康莱特注射液对肝癌化疗增敏作用的实验研究

目的 :研究康莱特 (KLT)注射液对肝癌细胞化疗增敏作用。方法 :用癌细胞药物敏感性试验(MTT法 )测定KLT注射液和 5种化疗药联合作用的敏感度与对照组比较。结果 :KLT注射液联合化疗药的抑瘤率高于单纯化疗药 (P <0 0 5)。结论 :KLT注射液分别和 5 FU、CP、DDP、MMC联用比单纯化学药物治疗肝癌有明显的增敏作用。     

康莱特联合化疗药物增敏的最佳时机的实验研究

目的:寻找康莱特联合化疗药物健择和顺铂的最佳时机。方法:采用MTT比色的方法。结果:康莱特对人肺腺癌95D的生长有抑制作用和化疗增敏作用;康莱特先于健择,顺铂加入对95D的抑制最强。结论:康莱特先于健择或顺铂加入优于两药同时加入。     

康莱特增加肺癌细胞对健择敏感性的实验研究

目的：研究康莱特联合化疗药物健择抑制人肺腺癌细胞95D生长的作用,同时寻找康莱特联合化疗药物健择作用的最佳时机。方法：采用MTT比色法进行检测。结果：康莱特单药对人肺腺癌95D的生长有抑制作用,联合健择的抑制率高于单药组;康莱特先于健择加入对95D的抑制最强。结论：康莱特体外对肺癌细胞有抗肿瘤和化疗增敏作用;康莱特先于健择加入对95D的抑制最强。     

大鼠肝癌细胞与间充质干细胞药敏相关性研究

背景与目的：探索针对肿瘤干细胞的药物敏感实验,对提高肝癌等实体瘤化疗有效率意义重大。在肝癌中,来自于骨髓干细胞的卵圆细胞是对应的成体干细胞。鉴于目前肿瘤干细胞和卵圆细胞分离困难,我们研究肝癌大鼠间充质干细胞与肿瘤细胞药敏相关性,探讨利用间充质干细胞代替肝癌肿瘤干细胞进行药物敏感实验的可能性。方法：以二乙基亚硝胺为诱癌剂制造大鼠肝癌模型。分离培养间充质干细胞与肝癌细胞,MTT法检测阿霉素（0．2μg／ml）、氟尿嘧啶（5μg／ml）及顺铂（1μg／ml）作用24h对细胞抑制率。化疗药物作用后的肝癌细胞移植裸鼠．6周后测量移植瘤重量。阿霉素、氟尿嘧啶及顺铂对这两种细胞抑制率以及对应药物作用后肝癌细胞移植瘤重量间进行相关性分析。结果：①3种药物对肝癌细胞抑制率与其作用肝癌细胞后移植瘤重量间进行相关性分析．具体偏相关系数为：阿霉素,0．6307（P〉0．05）;氟尿嘧啶,0．4358（P〉0．05）;顺铂,0．7080（P〉0．05）;②3种常用肝癌化疗药物对肝癌大鼠间充质干细胞抑制率与对肝癌细胞间抑制率进行相关性分析,具体偏相关系数为：阿霉素,0．6316（P〉0．05）;氟尿嘧啶,0．4214（P〉0．05）;顺铂,0．5943（P〉0．05）;药物对大鼠间充质干细胞抑制率与对应药物作用后肝癌细胞移植瘤重量有较好负相关性,具体偏相关系数为：阿霉素,-0．8308（P〈0．05）;氟尿嘧啶,-0．8991（P〈0．01）;顺铂,-0．8311（P〈0．05）。结论：化疗药物对肝癌细胞抑制率与对应药物作用后肝癌细胞移植瘤重量无相关性．常规药物敏感实验不能完全反映肝癌增殖侵袭能力。化疗药物对间充质干细胞抑制率与对应药物作用后肝癌细胞移植瘤重量有负相关,可以较好地反映肝癌细胞增殖侵袭能力。     

康莱特联合顺铂热灌注化疗治疗恶性腹腔积液的疗效观察

目的：探讨康莱特注射液联合顺铂热灌注化疗治疗恶性腹腔积液的疗效。方法：61例病人随机分为治疗组及对照组。治疗组31例采用康莱特注射液联合顺铂使用体腔热灌注机腹腔热灌注化疗;对照组30例单纯采用顺铂使用体腔热灌注机腹腔热灌注化疗。观察两组患者治疗前后实验室检查的变化、近期疗效、生存期及药物不良反应。结果：治疗组症状体征及实验室检查明显改善,近期疗效、生存期、药物不良反应均优于对照组（P〈0.05）。结论：应用康莱特联合顺铂热灌注化疗治疗恶性腹腔积液有一定的效果,值得推广应用。     

口腔颌面癌肿化疗药物敏感性检测对临床化疗的指导价值

目的本研究目的是评价口腔颌面癌肿活检标本体外化疗药敏检测结果对临床化疗效果和个体化疗方案制定的指导意义.方法 63例口腔颌面肿瘤病人纳入研究;化疗前活检切取肿瘤标本,用改良MTT法进行8种常用化疗药物顺铂(cisplatin,CDDP)、表阿霉素(epi-adriamycin,E-ADM)、长春地辛(vindesin,VDS)、5-氟尿嘧啶(5-fluorouracil,5-FU)、平阳霉素(pingyangmycin,PYM)、氨甲喋呤(methortrexatum,MTX)和紫杉醇(paclitaxel,Taxol).、替尼泊甙(teniposide,Vm-26)的药敏检测,63例病人接受不同方案的化学治疗,并对化疗近期疗效进行了评价.结果应用各药物的5倍峰血浆浓度得到8种药物对63例标本肿瘤细胞体外生长抑制率.化疗方案所用药物对肿瘤细胞的平均生长抑制率超过50%的有16例病人,临床化疗结果全部有效;平均生长抑制率超过30%而＜50%的有27例病人,临床化疗22例有效;而生长抑制率＜30%的20例病人,化疗结果仅7例有效.结论化疗药物对肿瘤细胞的体外平均生长抑制率和临床化疗有效率呈明显正相关,采用改良MTT法完成的化疗药敏检测对临床选择有效的化疗药物具有指导意义,提示体外药敏检测可用于口腔颌面癌肿的"个体化"化疗方案设计.     

康莱特增加肺癌细胞对健择敏感性的实验研究

目的:研究康莱特联合化疗药物健择抑制人肺腺癌细胞95D生长的作用,同时寻找康莱特联合化疗药物健择作用的最佳时机。方法:采用MTT比色法进行检测。结果:康莱特单药对人肺腺癌95D的生长有抑制作用,联合健择的抑制率高于单药组;康莱特先于健择加入对95D的抑制最强。结论:康莱特体外对肺癌细胞有抗肿瘤和化疗增敏作用;康莱特先于健择加入对95D的抑制最强。     

MTT比色分析法快速测定肝癌细胞株地化疗药物敏感性的方法研究

选用了人肝癌ＳＭＭＣ－７７２１细胞株对ＭＴＴ比色分析法用于体外抗肝癌化疗药敏试验中的几个变量进行了研究讨论，结果显示：ＭＴＴ甲产物最大吸收波长为５７０ｎｍ；ＭＴＴ用量以每孔２０μｌ（５ｍｇ／ｍｌ）为最宜；ＭＴＴ孵育６小时所得光吸收度（Ａ）值最高，甲量与细胞密度有好的线性关系，四种化疗药（阿霉素，卡铂，５－氟尿嘧啶，丝裂霉素）的作用时间以７２～９６小时为最好，且药物对细胞株的抑制率随药物浓度的增加而     

康莱特在原发性肝癌多学科综合治疗中的作用

目的：中晚期原发性肝癌患采用康莱特静滴联合肝动脉栓塞化疗，观察其在原发性肝癌综合治疗中的作用，方法：对住院确诊的中晚期原发性肝癌患100例，全部经皮穿刺股动脉，在X线透视下，将导管插至固有动脉及其分支。注射混合的阿霉毒、顺铂、羟基喜树碱各30mg和磺化油5－10ml，一般3－4周1次共3次，其中50例同应用康莱特注射液100ml，每日一次，连续21天，共2个周期，结果：单纯肝动脉栓塞化疗和中西医结合治疗在减轻癌性疼痛，肝功能损害恢复和缓解粒细胞减少程度，治疗前后有显性差异，瘤体缩小消失率，单纯肝动脉栓塞化疗组12％（6／50）远低于配合康莱特治疗的34％（17／50），P＜0．01）。结论：中西药物结合治疗肝癌，增效减毒，与单纯肝动脉栓塞化疗比较有显性差异。     

康莱特注射液对吉非替尼诱导人肺腺癌A549细胞株凋亡影响的实验研究

目的 探讨康莱特注射液联合吉非替尼对人肺腺癌A549细胞株增殖和凋亡的影响。 方法 应用MTT法计算康莱特注射液作用于人肺腺癌A549细胞株48 h的半数抑制浓度（IC50）,检测康莱特注射液（5、10、20、40、80、100、160 μl／ml）、吉非替尼（0.1、0.5、1.0、5、10、20、40 μmol／L）以及康莱特注射液（80 μl／ml）联合吉非替尼（0.1、0.5、1、5、10、20、40 μmol／L）作用于人肺腺癌A549细胞株24、48、72 h的增殖抑制率;流式细胞仪Annexin V/PI 双标法检测康莱特注射液、吉非替尼及两药联合作用于A549细胞48 h后的凋亡率;免疫细胞化学法检测各药物处理组作用于A549细胞48 h后FAS、AKT2蛋白表达;RT-PCR技术检测各组FAS mRNA、AKT2 mRNA的表达水平。均设未加药的A549细胞为空白对照组。结果 康莱特注射液作用于人肺腺癌A549细胞株48 h的IC50为80 μl／ml。康莱特注射液联合吉非替尼作用于A549细胞的增殖抑制率高于两单药组和空白对照组,且呈时间和浓度依赖性（P＜0.05）。两者的联合作用在一定浓度范围内呈现出协同的抗瘤效果。流式细胞术检测显示,两药联合组的细胞凋亡率显著高于两单药组和空白对照组（P＜0.05）。免疫细胞化学和RT-PCR检测显示,与空白对照组比较,康莱特组和两药联合组FAS蛋白和mRNA的表达水平均升高（P＜0.05）,AKT2蛋白和mRNA表达水平均下调（P＜0.05）。结论 康莱特注射液联合吉非替尼对人肺腺癌A549细胞株有明显的促凋亡作用,康莱特注射液可能增加人肺腺癌A549细胞对于吉非替尼的敏感性。     

康莱特诱导肿瘤细胞凋亡的实验研究

目的：探讨康莱特的肿瘤机制，特别是凋亡的作用机制。方法：应用形态学方法、流式细胞术、DNA凝胶电泳、Annexin V标记等方法检测细胞凋亡的发生。应用RT－PCR检测调相关基因Fas、bcl-2、c-myc表达的变化。结果：康莱特对人肺腺癌细胞A549和人早幼粒细胞性白血病细胞HL60的生长均有抑制作用，引起细胞死亡。康莱特对A549细胞诱导凋亡的反应并不明显，而对HL60细胞有明显的凋亡。在HL60凋亡过程中，凋亡相关基因Fas转录水平比用药前增强，c-myc和bc1-2含量无变化。结论：凋亡与坏死同为康莱特抑瘤的机制，但不同细胞系凋亡所发生的比例不同。提示不同肿瘤，其凋亡规律亦不同，为康莱特的临床应用提供了理论依据。     

Chinese herb related molecules of cancer-cell-apoptosis: a minireview of progress between Kanglaite injection and related genes

Many kinds of Chinese herb had been confirmed to have the character of anti-tumor, clinical reports about anti-tumor effects of Chinese herb had also been found in recent years, but most of the reports were focused on the clinical treatment of effectiveness for Chinese herb, on the other hand, review about Chinese herbal related with molecules on cancer-cell-apoptosis was seldom, many scientists could not believe such kinds of clinical describes about anti-tumor effects for Chinese herb, because these describes were lack of molecular biology evidence. Kanglaite(KLT) injection is an anti-tumor new drug which extracts from Chinese medicine-coix seed with modern advanced pharmaceutical technology, it is also a new biphase extended-spectrum anticancer medicine, the food and drug administration(FDA) of United States also approved a phase II trial of KLT to test its efficacy in treating non-small-cell lung cancer. Some studies show it could inhibit some anti-apoptotic gene and activate some pro-apoptotic gene, its injection solution is one of the new anticancer medicine that can significantly inhibit a various kinds of tumor cells, so it has become the core of research that how to further explore KLT injection to promote tumor cell apoptosis by impacting on related genes. In this review, the relationship between KLT and some tumor cell apoptosis molecules had been discussed and reviewed generally.     

Effects of kanglaite capsules combined with transcatheter arterial chemoembolization （TACE） on patients with mid or late-stage primary hepatocellular carcinoma （HCC）

Objective:To observe the clinical effects of kanglaite (KLT)capsules combined with transcatheter arterial chemo- embolization(TACE)in treating patients with mid or late-stage primary hepatocellular carcinoma (HCC).Methods:Sixty-five cases were randomly divided into 2 groups.32 patients in combination group received the treatment of KLT capsules+TACE and 33 patients in control group were treared with TACE alone.The objective response rate(RR).serum alpha fetoprotein (AFP),peripheral blood T lymphocyte subgroups(T-LS),quality of life(QOL),time to progression(TTP)and adverse reaction were observed and compared between 2 groups.Results:The objective response rate and serum alpha fetoprotein levels had no significant difference between the two groups (P＞0.05).Combination group was superior to control group in quality of life(QOL).time to progression(TTP),peripheral blood T lymphocyte subgroups(CD3+,CD4+,CD4+＼CD8 ratio)and liver adverse reactions,with significant differences (P＜0.05).Conclusion:KLT capsules combined with TACE is an effective method to treat primary hepatocellular carcinoma (HCC) patients who have lost the opportunity of surgical therapy.     

薏苡仁油注射液对人体肝癌SMMC-7721细胞株体外抗肿瘤作用及机制研究

目的：研究薏苡仁油注射液（KLT）对人体肝癌SMMC-7721的体外抗肿瘤作用及机制。方法：在人肝癌SMMC-7721细胞模型上采用CCK-8细胞增殖试验、划痕试验、Transwell小室穿膜试验、Matrigel克隆形成试验观察KLT对细胞增殖、迁移及侵袭的影响;应用流式细胞术检测KLT对肿瘤细胞周期及细胞凋亡的影响;Western blot检测KLT对肿瘤细胞中目的基因的表达情况。结果：经KLT作用后的人肝癌细胞生长、迁移及侵袭功能被抑制;流式细胞术检测发现KLT处理的肝癌细胞阻滞于G2/M期,细胞晚期凋亡较明显;KLT上调了cyclin B1的表达,下调了cyclin D1、cyclin E的表达。结论：KLT在体外对肝癌细胞具有良好的抗肿瘤活性,其作用机制可能与其诱导的细胞周期阻滞、细胞凋亡、抑癌基因的上调、癌基因的下调有关。     

Inhibition of HIF-1α by PX-478 enhances the anti-tumor effect of gemcitabine by inducing immunogenic cell death in pancreatic ductal adenocarcinoma

Pancreatic ductal adenocarcinoma (PDAC) is the worst prognoses among all the malignancies. Now, gemcitabine (Gem) is the first line chemotherapeutic drug for advanced pancreatic cancer. However, Gem is usually ineffective to the PDAC because of high degree of drug resistance. Hypoxia and immune suppressive milieu are the best-described hallmarks of PDAC; therefore, we investigated the impact of hypoxia inducible factor-1 (HIF-1) inhibitor, PX-478, in combination with Gem on the induction of immunogenic cell death (ICD). We verified that combined treatment with Gem/PX-478 significantly enhanced the anti-tumor effect and increased proportion of tumor infiltrating T-lymphocytes in Panc02-bearing immune-competent but not in immune-deficient mice. Vaccination using Panc02 cell line treated with single agent or in combination showed significant anti-tumor effects. Pancreatic cell lines treated with Gem and PX-478 can induce an increase in eIF2α phosphorylation was correlated with down-regulation of HIF-1α and elicited exposure of CRT and release of HMGB1 and ATP. Only co-treated cells induced DC maturation/phagocytosis and IFN-γ secretion by cytotoxic T lymphocytes. Altogether, combined treatment with Gem/PX-478 showed significantly inhibition on tumor growth and anti-tumor immunization. We propose that inhibition HIF-1α elicits Gem-induced immune response and eliminates PDAC cells by inducing ICD.     

Anti-tumor effects of tumor necrosis factor in combination with chemotherapeutic agents

The anti-tumor activity of partially purified tumor necrosis factor (TNF) was analysed in combination with chemotherapeutic drugs against intradermally transplanted Meth A sarcoma. Tumors were allowed to grow until they reached an average diameter of about 8 mm. TNF was given once i.v. (16,000 units as determined on L-M cells) and the chemotherapeutic agents adriamycin (1-10 mg/kg), 5-FU (3-100 mg/kg), cyclophosphamide (50-200 mg/kg) were applied once i.p. 4 hr after TNF injection. The strongest anti-tumor effects in normal BALB/c mice were observed when TNF was combined with the following doses of chemotherapeutic agents: cyclophosphamide 100 mg/kg, adriamycin 5 mg/kg, 5-FU 30-100 mg/kg. The most effective combinations induced complete regressions. When TNF was combined with varying doses of adriamycin and cyclophosphamide, bell-shaped dose-response curves were obtained. Experiments were repeated in Meth A sarcoma-bearing BALB/c nu/nu mice. In this case TNF combinations with cyclophosphamide and 5-FU did not induce tumor regressions. The highest dose of the chemotherapeutic agent was the most effective in drug combinations. Histological analysis revealed a potentiation of the TNF-induced necrosis by cyclophosphamide. Increased hyperemia and extravasation of erythrocytes could be found in tumors of animals treated with the drug combination.     

Gemcitabine has significant immunomodulatory activity in murine tumor models independent of its cytotoxic effects

Given reports that the chemotherapeutic agent gemcitabine (GEM) does not block T-lymphocyte recall responses and is not detrimental to specific anti-tumor immunity, studies to evaluate the use of GEM in combination with immunotherapy were initiated. When we tested the therapeutic effects of GEM as a single agent in various murine tumor models, we found that a single dose of GEM had impressive anti-tumor activity in a specific subset of tumors. Surprisingly, efficacy was not related to in vitro drug sensitivity, but instead, correlated with the immunogenicity of the tumor. A key role of the immune system in GEM's action was demonstrated in experiments showing that the anti-tumor effects of GEM were lost in nude mice. In addition, we saw equivalent anti-tumor effects of GEM in animals bearing tumors that were extremely resistant to the in vitro cytotoxic effects of GEM versus parental GEM-sensitive cells. This therapeutic efficacy was thus not due to direct cytotoxic effect on tumor cells, but rather to an enhancement of T-cell mediated anti-tumor immune effects. These data raise the exciting possibility that GEM may be a useful agent in combination with various types of tumor immunotherapy.