Human bone marrow mesenchymal stem cell-derived exosomes loaded with gemcitabine inhibit pancreatic cancer cell proliferation by enhancing apoptosis

BACKGROUND Pancreatic cancer remains one of the most lethal malignancies,and has limited effective treatment.Gemcitabine(GEM),a chemotherapeutic agent,is commonly used for clinical treatment of pancreatic cancer,but it has characteristics of low drug delivery efficiency and significant side effects.The study tested the hypothesis that human bone marrow mesenchymal stem cell(MSC)-derived exosomes loaded with GEM(Exo-GEM)would have a higher cytotoxicity against human pancreatic cancer cells by enhancing their apoptosis.AIM To investigate the cytotoxicity of MSC-derived Exo-GEM against pancreatic cancer cells in vitro.METHODS Exosomes were isolated from MSCs and characterized by transmission electron microscopy and nanoparticle tracking analysis.Exo-GEM through electroporation,sonication,or incubation,and the loading efficiency was evaluated.The cytotoxicity of Exo-GEM or GEM alone against human pancreatic cancer Panc-1 and MiaPaca-2 cells was assessed by MTT and flow cytometry assays.RESULTS The isolated exosomes had an average size of 76.7 nm.The encapsulation efficacy and loading efficiency of GEM by electroporation and sonication were similar and significantly better than incubation.The cytotoxicity of Exo-GEM against pancreatic cancer cells was stronger than free GEM and treatment with 0.02μM Exo-GEM significantly reduced the viability of both Panc-1 and MiaPaca-2 cells.Moreover,Exo-GEM enhanced the frequency of GEMinduced apoptosis in both cell lines.CONCLUSION Human bone marrow MSC-derived Exo-GEM have a potent cytotoxicity against human pancreatic cancer cells by enhancing their apoptosis,offering a promising drug delivery system for improving therapeutic outcomes.

Gemcitabine治疗晚期非小细胞肺癌的临床研究

目的：评价Ｇｅｍｃｉｔａｂｉｎｅ（中文译名：健择）单药及其与顺氯铵铂联合化疗方案治疗ＩＩ、ＩＶ期非小细胞肺癌（ＮＳＣＬＣ）的临床疗效及其不良反应。材料与方法：１健择单药研究：从１９９７年９月至１９９８年５月入选ＩＩ、ＩＶ期ＮＳＣＬＣ病人２１例，以前未接受过任何化疗、放疗。ＩＩ期病人１１例，ＩＶ期病人１０例。健择１０００ｍｇ／ｍ２，第１，８，１５天各注一次，每２８天为一疗程。２健择与顺氯铵铂联合化疗研究：从１９９７年１０月至１９９８年７月入选ＩＩ、ＩＶ期ＮＳＣＬＣ病人４８例，以前未接受过任何化疗、放疗。ＩＩ期病人１９例，ＩＶ期病人２９例。健择１０００ｍｇ／ｍ２，第１，８，１５天各注一次；顺氯铵铂１００ｍｇ／ｍ２，第一天用，每２８天为一疗程。结果：１健择单药研究：可评价疗效的有１９例，６例获部分缓解（ＰＲ），其中２例（１０５％）经４周以上复查证实。总有效率３１５％［９５％ＣＩ，９４％～４５１５％］。全组均可评价不良反应，少数病人发生轻微胃肠道反应、白细胞下降、血红蛋白下降和血小板下降，只有４８％（各１例）患者发生ＩＩ度的恶心呕吐及白细胞下降，有１例患者发生ＩＩ度感染。全组中位生存期６１３月。２?

microRNA-218 promotes gemcitabine sensitivity in human pancreatic cancer cells by regulating HMGB1 expression

Objective： The purpose of this study was to examine the effect of gemcitabine（GEM） on micro RNA-218（mi R-218） expression in human pancreatic cancer cells.Methods： Quantitative reverse transcription polymerase chain reaction（q RT-PCR） was performed to examine the differences in mi R-218 expression between the GEM-sensitive Bx PC-3 pancreatic cancer cells and GEM-resistant PANC-1 cells. The effect of GEM on the expression of mi R-218 in PANC-1 cells was also investigated. PANC-1 cells were transfected either with HMGB1 si RNA to knock down the expression of HMGB1 or with the recombinant HMGB1 expression vector（pc DNA3.1-HMGB1） to overexpress HMGB1. The effect of ectopic expression of HMGB1 on the apoptosis of mi R-218-transfected and GEMtreated PANC-1 cells was examined by flow cytometric analysis.Results： The mi R-218 expression level was lower in GEM-resistant PANC-1 cells compared to GEMsensitive Bx PC-3 cells（P〈0.05）. The percentage of apoptotic PANC-1 cells was significantly increased in the mi R-218 mimic ＋ GEM group compared to the mimic ctrl ＋ GEM group and the normal control group（P〈0.01）. The HMGB1 expression level was markedly decreased in PANC-1 cells transfected with HMGB1 si RNA but was significantly increased in PANC-1 cells transfected with the recombinant HMGB1 expression vector, pc DNA3.1-HMGB1（P〈0.01）. The proportion of apoptotic PANC-1 cells was significantly lower in the mi R-218 mimic ＋ GEM ＋ pc DNA3.1-HMGB1 group compared to the mi R-218 mimic ＋ GEM ＋ HMGB1 si RNA group（P〈0.01）.Conclusions： The expression level of mi R-218 was downregulated in the GEM-resistant cell line. mi R-218 promoted the sensitivity of PANC-1 cells to GEM, which was achieved mainly through regulating the expression of HMGB1 in PANC-1 cells.

ERK signaling pathway may induce gemcitabine chemoresistance in pancreatic cancer cell line SW1990 by regulating the expression of mdr-1 and RRM1 gene

Objective： To investigate the relationship between extracellular signal-regulated kinase （ERK） pathway, multidrug resistance gene （mdr-1）, ribonucleotide recluctase M1 （RRM1） gene and their roles in gemcitabine （GEM） chemoresistance in pancreatic cancer cell line SW1990. Methods： The GEM-resistance cell model was constructed by a stepwise method. Immunohistochemistry was used to measure the expression of ERK protein （ERK1/2） in the established cell strains in a semiquantitative way. The mRNA expression of mdr-1 and RRM1 were detected by RT-PCR. MTT assay was performed to determine the IC50 value. Results： The established GEM-resistant cell strains were able to gain stable growth and passage ability in the medium contained different concentration levels of GEM （0, 30, 60, 100, 150 and 200 nmol/L）. The expression of ERK protein, mdr-1 and RRM1 gene were elevated accompanied by the increase of GEM concentration. There was a highly positive correlation between mdr-1, RRM1 expression and GEM-resistanca level （r = 0.960, P = 0.002 and r = 0.966, P = 0.002）. The expression of ERK protein also correlated with the mdr-1 and RRM1 level （r = -0.943, P = 0.005 and r = -0.883, P = 0.02）. At the GEM-resistance level of 200 nmol/L, the grey scale value of ERK1/2 was 164.22 ±13.17, mdr-1/β-actin and RRM1/β-actin were 1.41 ±0.04 and 1.45 ± 0.18, respectively; after treated with ERK pathway inhibitor U0126, these values synchronously decreased to 186.85 ± 13.14, 0.2 3± 0.02 and 0.21 ± 0.03, respectively; inversely, the ERK1/2 grey scale value was 106.55 ± 16.45, mdr-l/β-actin and RRMl/β-actin were 1.50± 0.07 and 1.52 ± 0.12, respectively, which presented a tendency of synchronously increase after treated with ERK pathway activator EGF. The IC50 values in GEM-resistant cells of 0 nmol/L and 200 nmol/L levels were 4.104 and 10.20, respectively. After treated with U0126, these values decreased to 3.26 and 4.50, respectively; while treated with EGF, the IC50 values increased to 8.89 and 17.17, respectively. Conclusion： The ERK pathway may induce the GEM-chemoresistance in pancreatic cell line SW1990 through the participation in the regulation of the mdr-1 and RRM1 gene expression.

过表达Plk-1对gemcitabine诱导的胰腺癌细胞化疗敏感性的影响

目的观察Plk-1基因过表达对gemcitabine诱导胰腺癌细胞AsPC-1化疗敏感性的影响。方法①将携带有人外源性Plk-1基因的真核表达质粒转染AsPC-1细胞系设为处理组,转染空载体设为对照组,无处理组设为空白组,转染24 h后,提取细胞总RNA和总蛋白,利用RT-PCR法及Western blot鉴定AsPC-1细胞内Plk-1基因和蛋白表达水平。②采用流式细胞术检测转染组细胞凋亡的改变。③不同浓度gemcitabine作用于转染细胞,MTT法测定转染48 h后细胞增殖能力。结果①测序结果表明成功的从AsPC-1细胞总RNA中扩增Plk-1基因。RT-PCR结果表明:未转染组AsPC-1细胞组、转染空载体pcDNA3.1组及pcDNA3.1/Plk-1组24 h各组细胞内Plk-1 mRNA相对量分别为(2.14±0.16)、(2.18±0.15)、(2.58±0.18),转染pcDNA3.1/Plk-1组与其他各组相比,差异有高度统计学意义(P<0.01)。Western blot结果显示:未转染组AsPC-1细胞组、转染空载体pcDNA3.1及pcDNA3.1/Plk-1组24 h各组细胞内Plk-1基因的蛋白表达水平为(0.989±0.018)、(1.022±0.021)、(1.243±0.143),转染pcDNA3.1/Plk-1组与其他各组相比,差异有高度统计学意义(P<0.01)。②在gemcitabine作用下,pcDNA3.1/Plk-1转染组细胞凋亡率明显低于未转染组及空载体转染组,差异有高度统计学意义(P<0.01)。③AsPC-1/Plk-1转染组细胞的生长抑制率亦明显高于未转染组及空载体转染组,差异有高度统计学意义(P<0.01)。结论胞浆过表达Plk-1活性分子可明显降低gemcitabine诱导的AsPC-1细胞凋亡,增强其对化疗药物的耐受性。

吉西他滨(Gemcitabine)联合铂类药物治疗老年中晚期非小细胞肺癌(NSCLC)的临床观察

目的 :探讨吉西他滨 (Gemcitabine)联合铂类药物治疗老年中晚期非小细胞肺癌 (NSCLC)的临床疗效和毒性。方法 :予吉西他滨 (Gemcitabine) 10 0 0mg/m2 第 1、 8天静滴并联合铂类药物治疗老年非小细胞肺癌(NSCLC) 2 0例 ,三周为一治疗周期 ,两周期后评价疗效和毒副作用 ,随访缓解期和生存期。结果 :本组 2 0例病例中 ,治疗总有效 4 0 % ,中位缓解期 7个月 ,中位生存期 9个月 ,主要毒副作用为骨髓抑制和胃肠道反应。其中白细胞减少发生率为 80 % ,血小板减少发生率为 6 0 %。结论 :吉西他滨 (Gemcitabine)对老年非小细胞肺癌(NSCLC)有较好疗效 ,主要毒性反应为较严重的血液学毒性 ,老年患者最好辅以G -CSF或GM -CSF以及免疫与营养支持治疗 ,则能顺利完成化疗 ,吉西他滨 (Gemcitabine)联合铂类药物可做为老年非小细胞肺癌 (NSCLC)的一线治疗方案。

Treatment of Primary Liver Cancer with Postoperative After-Loading Radiotherapy and Gemcitabine

Objective： To evaluate the curative effectiveness of postoperative after-loading radiotherapy with the use of gemcitabine in 22 patients with primary liver cancer. Methods： From Oct. 1999 to Dec. 2001, 22 patients with primary liver cancer underwent postoperative after-loading radiotherapy 3-10 days after hepatectomy and chemotherapy using gemcitabine （1400 mg every week for 3 weeks, repeated after one week interval, total cycles were 6） and compared with 22 cases of sole hepatectomy. Three-six catheters were placed for irradiation after hepatectomy. The single-dose of after-loading radiotherapy was 10 Gy, 24 sessions per person. Results＇. The rate of AFP negative-reversion was 100% （17/17） in the treated group, higher than in control group （62.5%, 10/16, P〈0.05）. In the treated group, the 1-year relapse rate, metastasis rate and survival rate were 18.2% （4/22）, 0 and 100% （22/22） respectively, while in the control group they were 45.5% （10/22）, 13.6% （3/22） and 77.3% （17/22） respectively. There were significant differences between the two groups in relapse rate, metastasis rate and survival rate within a year （P〈0.05）. Conclusion： Postoperative after-loading radiotherapy with gemcitabine is an effective way for the treatment of primary liver cancer.

Experimental study of gemcitabine combination with radiation in vitro on a highly metastatic human ovarian cancer cell line HO-8910PM

Objective： The aim of the study was to investigate the mechanism of gemcitabine （GEM） combination with radiation on the high metastasis human ovarian cancer cell line （HO-8910PM）. Methods： Human ovarian cancer cell line HO- 8910PM was treated with different concentrations of gemcitabine for 24 h, then the cells were counted. In the study of GEM combination with radiation, an efficiency of colony formation was observed; the cell cycle and apoptosis were analyzed by flow cytometry; the experiment of depend on the time and its radio sensitivity were observed by using mitotic index with the cells for each 24, 48, 72 and 96 h after experiment. Results： It suggested that the GEM had an inhibition effect on the human ovarian cancer cell line. The alive cell numbers were decreased by following a height of GEM concentration. When GEM in combina- tion with a radiation, the suppression was significantly increased than that of single GEM therapy. The efficiency of colony formation was significantly lower, under this condition the cell could be arrested at G0-G1 phase and could be decreased to enter into the S phase; the apoptosis percentage could be significantly increased; especially, under the 4 Gy and 6 Gy doses the cell apoptosis was more obvious. GEM combination with radiation had depended on the time to the cells; mitotic index of the calls in combination group was observed significantly lower than that of single GEM therapy or single radiation, and this showed that it had an effect of radiosensitivity. Conclusion： The GEM has a significant growth inhibition on the human ovarian cancer cells, GEM combination with radiation could induce HO-8910PM cell occurred arrested and apoptosis. It has depended on the time and has a radiosensitivity effect. The result shows that it is a better method to treat the human ovarian cancer by using radiotherapy combined with gemcitabine.

Treatment of Unresectable Carcinoma of Pancreas with ^(125)I Implantation and ^(125)I Plus Gemcitabine

Objective: To study the role of 125 I and 125 I plus gemcitabine (GEM) in treatment of unresectable carcinoma of pancreas. Methods: From April 2000 to April 2003, 38 untreated patients with locally advanced pancreatic cancer (LAPC) were collected and randomized into two groups: Arm A 125 I (18 patients) and Arm B 125 I+GEM (20 patients). Eligibility criteria were: cytologically and pathologically proven pancreatic carcinoma, Karnofsky performance status (kps) 60 80, age 18 75 years, adequate hematological, renal and liver function, and controllable pain. Arm A patients were treated with 125 I implants. Arm B patients started chemotherapy within 10 14 d post operatively following the implant procedure. Chemotherapy doses were as follows: GEM 1 000 mg/m 2 weekly × 3 followed by 1 week of rest for 3 cycles. In addition, all patients underwent laparotomy and surgical staging. The surgical procedures performed were biopsy, gastric bypass and biliary bypass. The total activity and number of seeds used were as recommended by Anderson. The mean activity, minimal peripheral dose (MPD), and volume of implants were 20 mCi, 14 000 cGy, and 53 cm 3, respectively. Results: Overall response rate (CR+PR) in Arm A was 37.6% and in Arm B it was 44.5% ( P >0.05). PR median duration in Arm A was 6.7 months and in Arm B it was 4.8 months ( P <0.05). Clinical benefit response was experienced by 11.7 % of Arm A compared with 42.1% of Arm B ( P <0.05). The incidences of hematological toxicity (such as neutropenia) between Arm A and Arm B were 5.8% and 21.1%, respectively ( P >0.05). The survival rates of 12 and 24 month were 32.5%, 16.3% for Arm A and 61%, 38.7% for Arm B ( P =0.04). The rate of complication of Arm A was lower than that of Arm B without statistical significance. Conclusion: To some extent, 125 I or 125 I plus GEM is able to lead to a moderate objective response for LAPC with obstructive jaundice on the base of biliary bypass or/and gastric bypass, but 125 I plus GEM is more effective than 125 I in improvement of the quality of life and survival rate in patients with LAPC.

基因芯片技术筛选Gemcitabine诱导胰腺癌Panc-1细胞凋亡相关基因的研究

目的研究抗癌药吉斯他滨（Gemcitabine）诱导胰腺癌Panc-1细胞凋亡的相关基因。方法应用基因芯片技术检测50．33μg／mL Gemcitabine作用于胰腺癌Panc-1细胞48h后凋亡基因表达的变化。结果Gemcitabine抑制细胞表达14种凋亡相关基因，包括IRF-4、Scotin、14．3．3gamma、PDE4、GRPS、TID1、AFP、STAT5、Sox-4、c-Rel、MYCN、Htt、CatD、Gd。相反，gemcitabine促进了3种凋亡相关基因的表达，包括CyclinB1、ASK1、AKT2。结论Gemcitabine诱导胰腺癌Panc-1细胞凋亡通过非依赖caspase ASK1途径。

hENT1 expression is predictive of gemcitabine outcome in pancreatic cancer: A systematic review

High human equilibrative nucleoside transporter 1(hENT1)-expression has shown a survival benefit in pancreatic cancer patients treated with gemcitabine in several studies.The aim of this systematic review was to summarize the results and try to assess the predictive value of hENT1 for determining gemcitabine outcome in pancreatic cancer.Relevant articles were obtained from PubMed,Embase and Cochrane databases.Studies evaluating hENT1-expression in pancreatic tumor cells from patients treated with gemcitabine were selected.Outcome measures were overall survival,disease-free survival(DFS),toxicity and response rate.The database searches identified 10 studies that met the eligibility criteria,and a total of 855 patients were included.Nine of 10 studies showed a statistically significant longer overall survival in univariate analyses in patients with high hENT1-expression compared to those with low expression.In the 7 studies that reported DFS as an outcome measure,6 had statistically longer DFS in the high hENT1 groups.Both toxicity and response rate were reported in only 2 articles and it was therefore hard to draw any major conclusions.This review provides evidence that hENT1 is a predictive marker for pancreatic cancer patients treated with gemcitabine.Some limitations of the review have to be taken into consideration,the majority of the included studies had a retrospective design,and there was no standardized scoring protocol for hENT1-expression.

Propofol induces apoptosis and increases gemcitabine sensitivity in pancreatic cancer cells in vitro by inhibition of nuclear factor-κ B activity

AIM:To investigate the effect of propofol on human pancreatic cells and the molecular mechanism of propofol action.METHODS:We used the human pancreatic cancer cell line MIAPaCa-2 for in vitro studies measuring growth inhibition and degree of apoptotic cell death induced by propofol alone,gemcitabine alone,or propofol followed by gemcitabine.All experiments were conducted in triplicate and carried out on three or more separate occasions.Data were means of the three or more independent experiments±SE.Statistically significant differences were determined by two-tailed unpaired Student’s t test and defined as P<0.05.RESULTS:Pretreatment of cells with propofol for 24 h followed by gemcitabine resulted in 24%-75% growth inhibition compared with 6%-18%when gemcitabine was used alone.Overall growth inhibition was directly correlated with apoptotic cell death.We also showed that propofol potentiated gemcitabine-induced killing by downregulation of nuclear factor-κB(NF-κB).In contrast,NF-κB was upregulated when pancreatic cancer cells were exposed to gemcitabine alone,suggesting a potential mechanism of acquired chemoresistance.CONCLUSION:Inactivation of the NF-κB signaling pathway by propofol might abrogate gemcitabineinduced activation of NF-κB,resulting in chemosensitization of pancreatic tumors to gemcitabine.

Nedaplatin/Gemcitabine Versus Carboplatin/Gemcitabine in Treatment of Advanced Non-small Cell Lung Cancer: A Randomized Clinical Trial

Objective： To evaluate the efficacy and safety of nedaplatin/gemcitabine （NG） and carboplatin/gemcitabine （CG） in the management of untreated advanced non-small cell lung cancer （NSCLC）. Methods： Sixty-two patients with previously untreated advanced NSCLC were recruited between June 2006 and November 2007. Subjects were randomly assigned to the NG arm （n=30） and the CG arm （n=32）. Only patients （24 and 25 in the NG and CG arms, respectively） who completed 〉2 chemotherapy cycles were included in the data analysis. The primary outcome measure was the objective response rate （ORR）. The secondary outcome measures included progression-free survival （PFS）, overall survival （OS） and adverse events. Results： There were no statistically significant differences in the efficacy measures （ORR, P=0.305; median PFS, P=0.298, median OS, P=0.961） or in the major adverse events （grade 3/4 neutropenia, P=0.666; grade 3/4 anemia, P=0.263; grade 3/4 thrombocytopenia, P=0.222） between the two treatment arms. However, there was a trend towards higher ORR （37.5% vs. 24.0%）, longer PFS （6.0 vs. 5.0 months）, and less adverse events in the NG arm. Conclusion： NG regimen seems to be superior over CG regimen for advance NSCLS, but further investigation is needed to validate this superiority.

Neoadjuvant sorafenib combined with gemcitabine plus oxaliplatin in advanced hepatocellular carcinoma

This paper reports the first case of a patient with hepatocellular carcinoma with lymph node metastasis treated by sorafenib combined with gemcitabine plus oxaliplatin,with a partial response and normalization of α fetoprotein,which allowed curative surgery.The potential synergy between these three drugs needs to be confirmed,and is currently being investigated in a randomized phase Ⅱ trial.

Weekly albumin-bound paclitaxel/cisplatin versus gemcitabine/cisplatin as first-line therapy for patients with advanced non-small-cell lung cancer:A phase II open-label clinical study

Objective: The aim of this trial was to compare both the efficacy and the safety of a weekly nanoparticle albumin-bound paclitaxel(nab-paclitaxel) plus cisplatin vs. gemcitabine plus cisplatin in patients with advanced non-small-cell lung cancer(NSCLC).Methods: A total of 84 participants received either 100 mg/m^2 nab-paclitaxel each week on d 1, 8 and 15 of a 28 day cycle, as well as cisplatin 75 mg/m^2 on d 1 every three weeks(nab-TP arm); or gemcitabine 1,000 mg/m^2 on d 1 and 8, plus cisplatin 75 mg/m^2 on d 1 every three weeks(GP arm). The primary end point was progression-free survival(PFS). The secondary end points were overall response rate(ORR) and overall survival(OS).Results: According to our analysis, the median PFS was 4.8 months for the nab-TP arm vs. 5.2 months for the GP arm(P=0.55). Analysis showed the median OS was 14.6 months for participants who were in the nab-TP arm vs. 15.1 months for those in the GP arm(P=0.94). Besides, nab-TP showed OS advantages over GP in patients harboring epidermal growth factor receptor(EGFR) mutation(26.7 vs. 15.3 months, P=0.046) and patients with a performance status of 0(23.5 vs. 14.7 months, P=0.020). It was found that incidences of drug-related grade 3 or 4 toxicities were comparable between the two treatment arms.Conclusions: Therefore, it can be seen that weekly nab-TP treatment has a similar efficacy and tolerability to GP treatment for patients who are undergoing their first-line treatment for NSCLC. It could be that survival differences among platinum doublets in the context of both EGFR mutation and performance status have the potential to be the basis for our further clinical trials.

A phase Ⅱ study of gemcitabine and carboplatin combination chemotherapy in gallbladder carcinoma

BACKGROUND: Patients with carcinoma of the gallbladder have advanced, unresectable tumor at the time of presentation and face a dismal prognosis in the absence of a standard palliative chemotherapy regimen. This study was undertaken to evaluate the efficacy and safety of combined chemotherapy of gemcitabine and carboplatin in 20 patients with advanced gallbladder carcinoma. METHODS: The criteria of eligibility included chemonaive patients with unresectable gallbladder cancer, bidimensionaIly measurable disease, Zubrod's performance status≤2, and adequate major organ function. The patients received gemcitabine (1000 mg/m^2) on days 1 and 8, and carboplatin (target AUC of 5.0 mg/ml) on day 1, in a 21-day cycle. CT was used for response assessment. RESULTS: In this group of 20 patients with advanced gallbladder carcinoma 6 were men and 14 women, with a median age of 55 years. The stage of the tumor at presentation was IVB in 14 patients (70%), IVA in 3 (15%) and Ⅲ in 3 (15%). Four patients (21%) achieved a complete response, and 3 (15.7%), a partial response; an overall response rate was 36.7%. The median time to progression of the tumor was 33.8 weeks, and 1-year survival rate of the patients was 43.3%. Anemia of WHO grade Ⅲ or Ⅳ was seen in 2 patients (10%) and 1 patient (5%), respectively. Grade Ⅲ neutropenia and thrombocytopenia were observed in 2 patients (10%) and 1 patient (5%), respectively. CONCLUSION: With mild toxicity, combined chemotherapy of gemcitabine and carhoplatin is effective in the treatment of advanced gallbladder carcinoma.

Oncogenic ADAM28 induces gemcitabine resistance and predicts a poor prognosis in pancreatic cancer

BACKGROUND Pancreatic cancer is a major cause of cancer-related death,with a 5-year overall survival rate being below 5%.The main causes of poor prognosis in pancreatic cancer include easy metastasis,high recurrence rate,and robust drug resistance.Gemcitabine is a first-line drug for patients with unresectable pancreatic cancer.However,due to drug resistance,the clinical effect is not satisfactory.ADAM28 is reported as a tumor promoter in some cancers,but its role in pancreatic cancer and gemcitabine chemoresistance in pancreatic cancer has not been elucidated.AIM To identify if ADAM28 can act as an important target to reverse the gemcitabine drug resistance in pancreatic cancer.METHODS RNA-sequence analysis was applied to explore the potential targets involved in the gemcitabine of pancreatic cancer.SW1990 pancreatic cancer cells were treated with an increased dose of gemcitabine,and the mRNA levels of ADAM28 were evaluated by RT-PCR.The protein and mRNA levels of ADAM28 were confirmed in the gemcitabine resistant and parallel SW1990 cells.The ADAM28 expression was also assessed in TCGA and GEO databases,and the results were confirmed in the collected tumor and adjacent normal tissues.The overall survival(OS)rate and relapse-free survival(RFS)rate of pancreatic cancer patients with high ADAM28 level and low ADAM28 level in TCGA were evaluated with Kaplan-Meier Plotter.Furthermore,the OS rate was calculated in pancreatic cancer patients with high tumor mutation burden(TMB)and low TMB.CCK-8 assay was used to examine the effect of ADAM28 on the viability of SW1990 cells.The ADAM28 and its co-expressed genes were analyzed in the cBioPortal for cancer genomics and subjected to GSEA pathway analysis.The correlations of ADAM28 with GSTP1,ABCC1,GSTM4,and BCL2 were analyzed based on TCGA data on pancreatic cancer.RESULTS RNA-sequence analysis identified that ADAM28 was overexpressed in gemcitabine-resistant cells,and gemcitabine treatment could induce the expression of ADAM28.The mRNA and protein levels of ADAM28 were elevated in gemcitabine-resistant SW1990 cells compared with parallel cells.Also,the expression of ADAM28 was upregulated in pancreatic tumor tissues against normal pancreatic tissues.Notably,ADAM28 was highly expressed in the classical type than in the basal tumor type.Furthermore,the high expression of ADAM28 was associated with low OS and RFS rates.Interestingly,the high levels of ADAM28 was associated with a significantly lower OS rate in the high TMB patients,but not in the low TMB patients.Moreover,overexpression of ADAM28 could reduce the cell viability inhibition by gemcitabine,and knockdown of ADAM28 could enhance the proliferation inhibition by gemcitabine.The GSEA analysis showed that ADAM28 was related to the regulation of drug metabolism,and ADAM28 was significantly positively correlated with GSTP1,ABCC1,GSTM4,and BCL2.CONCLUSION This study demonstrates that ADAM28 is overexpressed in pancreatic cancer,and closely involved in the regulation of gemcitabine resistance.Overexpression of ADAM28 is a novel prognostic biomarker in pancreatic cancer.

Clinical efficacy of gemcitabine and cisplatin-based transcatheter arterial chemoembolization combined with radiotherapy in hilar cholangiocarcinoma

BACKGROUND Radical surgical resection is regarded as the best treatment for hepatic hilar cholangiocarcinoma. However, 60%-70% of patients have lost the chance of surgery at the time of diagnosis. Simple biliary stent or drainage tube placement may fail in a short time due to tumor invasion or overgrowth, bile accumulation, or biofilm formation. Effective palliative treatments to extend the effective drainage time are of great significance for improving the quality of life of patients and changing the prognosis of patients. AIM To investigate the clinical efficacy of gemcitabine and cisplatin-based transcatheter arterial chemoembolization (TACE) combined with radiotherapy in hilar cholangiocarcinoma.METHODS A retrospective analysis was conducted on patients clinically diagnosed with hilar cholangiocarcinoma from June 2014 to January 2017 at the Liaoning Provincial Cancer Hospital. Patients were evaluated by specialists, and those who were not suitable for surgery or unwilling to undergo surgery and met the inclusion criteria were included in the study. There were a total of 72 patients (34 males and 38 females) with an average age of 59.9 years (range, 40-72 years). According to percutaneous transhepatic biliary angiography and the patients’ wishes, stent implantation or biliary drainage tube implantation was used to relieve biliary obstruction. The patients were divided into either a control group or a combined treatment group according to their follow-up treatment. The control group consisted of a total of 35 patients who received simple biliary drainage tube placement and biliary stent implantation (7 patients with bilateral stents and 6 with a unilateral stent) and 22 patients receiving biliary drainage tube placement alone. The combined treatment group received TACE and extracorporeal radiotherapy after biliary drainage or biliary stent implantation and consisted of a total of 37 patients, including 21 patients receiving combined treatment after biliary stent placement (14 patients with bilateral stents and 7 with a unilateral stent) and 16 undergoing combined therapy after implanting the biliary drainage tube. In the combination treatment group, the TACE chemotherapy regimen employed gemcitabine and cisplatin, and the embolic agent was iodized oil. A particular dose was determined according to the patient's body surface area and the tumor staining indicated by DSA. In vitro radiotherapy was performed with intensity-modulated radiotherapy or threedimensional conformal radiotherapy at an average dose of 48.3 Gy. Both groups were followed from stent implantation or drainage tube implantation until the patient quitted or died. The median length of follow-up observation was 13 mo. The differences in overall survival time and the effect of different jaundice reducing methods (single stent, double stent, or biliary drainage) on the patency time and survival time of biliary stents were compared between the two groups;the related factors affecting overall survival time were analyzed. RESULTS The median survival time of the control group was 10.5 mo;the median survival time of patients with biliary stent implantation and those with percutaneous biliary drainage was 9.6 mo and 11.4 mo, respectively, and there was no statistically significant difference between them. The median survival time of the combined treatment group was 20.0 mo, which was significantly higher than that of the control group (P < 0.05). Among patients in the combined treatment group, the median survival time of patients who underwent biliary stent implantation and those who accepted percutaneous biliary drainage before the combination therapy was 19.5 mo and 20.1 mo, respectively, and there was no significant difference between them. In the combination treatment group, the mean time of median stent patency was 15.6 mo, which was significantly higher than that of the control group (7.0 mo;P < 0.05). The independent factors affecting survival time included age, whether to receive combination therapy, percutaneous biliary drainage tube implantation, and Bismuth-Corlette classification as type IV. CONCLUSION Gemcitabine and cisplatin-based TACE combined with radiotherapy can prolong the survival of patients with hilar cholangiocarcinoma. Independent predictors of survival include selection of combination therapy, Bismuth-Corlette classification as type IV, selection of percutaneous biliary drainage tube implantation, and age.

Folate-chitosan-gemcitabine core-shell nanoparticles targeted to pancreatic cancer

Objective：Human pancreatic cancer is one of the most common clinical malignancies.The effect of comprehensive treatment based on surgery is general.The effects of chemotherapy were not obvious mainly because of lack of targeting and chemoresistance in pancreatic cancer.This study aimed to investigate the effects of folate receptor （FR）-mediated gemcitabine FA-Chi-Gem nanoparticles with a core-shell structure by electrostatic spray on pancreatic cancer.Methods：In this study,the levels of expression of FR in six human pancreatic cancer cell lines were studied by immunohistochemical analysis.The uptake rate of isothiocyanate-labeled FA-Chi nanoparticles in FR high expression cell line COLO357 was assessed by fluorescence microscope and the inhibition rate of FA-Chi-Gem nanoparticles on COLO357 cells was evaluated by MTT assay.Moreover,the biodistribution of PEG-FA-ICGDER02-Chi in the orthotopic pancreatic tumor model was observed using near-infrared imaging and the human pancreatic cancer orthotopic xenografts were treated with different nanoparticles and normal saline control.Results：The expression of FR in COLO357 was the highest among the six pancreatic cancer cell lines.The FR mainly distributed on cell membrane and fewer in the cytoplasm in pancreatic cancer.Moreover,the absorption rate of the FA-Chi-Gem nanoparticles was more than the Chi nanoparticles without FA modified.The proliferation of COLO357 was significantly inhibited by FA-Chi-Gem nanoparticles.The PEG-FA-ICGDER02-Chi nanoparticles were enriched in tumor tissue in human pancreatic cancer xenografts,while non-targeted nanoparticles were mainly in normal liver tissue.PEG-FA-Gem-Chi significantly inhibited the growth of human pancreatic cancer xenografts （PEG-FA-Gem-Chi vs.Gem,t=22.950,P=0.000）.Conclusions：PEG-FA-FITC-Chi nanoparticles might be an effective targeted drug for treating human FR-positive pancreatic cancer.