Establishment and preliminary application of organoids in ovarian cancer / 中华妇产科杂志

Objective:To explore the establishment and application of ovarian cancer organoids.Methods:Fresh ovarian tumor tissues, obtaining from patients underwent surgery in the First Affiliated Hospital of Nanjing Medical University between October 2021 and March 2022, were collected, enzymatic degraded, digested, and embedded into matrigel to establish organoids. A total of 32 ovarian cancer samples were collected. Hematoxylin eosin (HE) staining and immunofluorescence (IF) procedure were used to verify the morphological structure of organoids and their expression of molecular markers. 3D cyto-live or dead assay was used to detecte the live or dead cells in organoids. Carboplatin with a concentration ranging from 5 to 80 μmol/L (5, 10, 20, 40, 80 μmol/L) was added to organoids to calculate the 50% inhibitory concentration (IC 50) in different organoids. Results:(1) Organoids from a total of 32 patients were established, of which 18 cases could be passaged stably in the long term in vitro, while 14 could be passaged in the short time. The average amplification time of long-term passage in vitro was over 3 months, and the longest reached 9 months. (2) In HE staining, significant nuclei atypia and local micropapillary structures were observed in organoids. IF staining revealed that ovarian cancer organoids expressed molecular markers similar to primary tumor tissues, such as Pan cytokeratin (Pan-CK), p53, paired box gene 8 (PAX8), and Wilms tumor gene 1 (WT1). (3) In 3D cyto-live or dead assay, a large number of apoptotic cells were observed inside and around the organoids after added carboplatin. The sensitivity to carboplatin varied in 18 organoids could amplify in the long term, with an average IC 50 of (29.5±15.8) μmol/L. Moreover, IC 50 values of 4 organoids derived from patients received neoadjuvant chemotherapy were much higher than the 14 organoids which did not received neoadjuvant chemotherapy [(48.7±11.3) μmol/L vs (24.0±12.1) μmol/L; t=3.429, P=0.022]. Conclusions:Organoids recapitulate ovarian cancers in vitro and could be stably passaged. Organoids derived from patients received neoadjuvant chemotherapy have higher resistance to carboplatin.

Correlation between neoadjuvant radiotherapy and chemotherapy efficacy and changes in peripheral blood T lymphocytes in patients with advanced mid-to-low rectal cancer / 中国基层医药

Objective:To correlate neoadjuvant radiotherapy and chemotherapy efficacy with changes in peripheral blood T lymphocytes in patients with advanced mid-to-low rectal cancer.Methods:A total of 106 patients with rectal cancer who received treatment in Jinhua Municipal Central Hospital from January 2019 to December 2020 were included in this study. Fasting venous blood was taken before neoadjuvant radiotherapy and chemotherapy and 7 days before surgery to measure the numbers of CD3 +, CD4 +, CD8 +, CD45RA + and CD45RO + cells using flow cytometry. The optimal cut-off point was determined using the receiver operating curve. The influential factors of tumor regression grade were analyzed using logistic regression analysis. Results:After neoadjuvant radiotherapy and chemotherapy, the numbers of CD3 +, CD4 +, CD8 + cells were (401.86 ± 138.65), (225.83 ± 87.17), and (155.84 ± 71.19) respectively, which were significantly decreased compared with before neoadjuvant radiotherapy and chemotherapy [(477.33 ± 141.74), (647.38 ± 203.19), (348.22 ± 113.75), t = 10.78, 11.17, 9.49, all P < 0.05]. There were no significant differences in the percentages of CD3 +, CD4 +, CD8 +, CD45RA + and CD45RO + cells between before and after treatment (all P > 0.05). The percentage of CD45RO + cells was significantly increased after neoadjuvant radiotherapy and chemotherapy. A higher percentage of CD45RO + cells led to a lower tumor regression grade ( P < 0.05). The receiver operating characteristic curve showed that the optimal cut-off point of the percentage of CD45RO + cells was 1.08. The area under the receiver operating characteristic curve was 0.774 ( P = 0.029), with a sensitivity of 82.5% and specificity of 69.6%. The logistic regression analysis revealed that the percentage of CD45RO + cells was significantly correlated with tumor regression grade ( P < 0.05). Conclusion:The percentage of CD45RO + cells in T lymphocyte subsets before and after neoadjuvant radiotherapy and chemotherapy is closely related to tumor regression grade. It can be used as an indicator to predict the sensitivity of neoadjuvant radiotherapy and chemotherapy. This study is of great innovation and science and provides a new idea for clinical practice.

Hepatobiliary tumor models and novel technology for accurate drug screening / 临床肝胆病杂志

Hepatobiliary tumor is a type of malignant tumor including primary liver cancer, cholangiocarcinoma, and gallbladder carcinoma. At present, hepatobiliary tumors have become the second leading cause of cancer-related death worldwide, while the treatment methods for such tumors cannot effectively meet clinical needs. Therefore, it is a key scientific problem in this field to explore and develop the experimental technology of accurate drug screening for hepatobiliary tumors, find new strategies and methods for clinical treatment, and provide new ideas for early diagnosis and comprehensive treatment of hepatobiliary tumors. This article introduces the latest research advances in the novel technologies for accurate drug screening for hepatobiliary tumor and their application potential by focusing on the construction of individualized pathological models of hepatobiliary tumor, drug screening technologies, the design and screening strategy of specific target drugs, and drug screening strategy based on artificial intelligence and big data analysis, as well as the directions for future development.

Multi-omics molecular subgrouping of hepatocellular carcinoma and its application in precision diagnosis and treatment / 临床肝胆病杂志

Hepatocellular carcinoma (HCC) is the third leading cause of cancer mortality in China. In recent years, the application of targeted therapy and immunotherapy has improved the survival rate of HCC patients. However, a significant difference in treatment response is observed among HCC patients due to tumor heterogeneity and a lack of biomarkers to predict efficacy. The advance in proteogenomics-centered multi-omics studies and the development of high-throughput drug screening platforms will help to develop new clinical treatment strategies for HCC and new methods for predicting the efficacy of precision medication, thereby realizing personalized precision diagnosis and treatment.

Chemical constituents from the heartwood of Trachycarpus fortunei (Hook.) H. Wendl / 军事医学

Objective To study the chemical constituents of the heartwood of Trachycarpus fortunei.Methods Compounds were isolated and purified by silica gel column , Sephadex LH-20 chromatography and recrystallization .Their structures were elucidated by physicochemical properties and spectral data .Results Ten compounds were isolated from the 95%ethanol extract of the heartwood of T.fortunei and identified as 1,3-dihydroxyanthraquinone (1),mollugin(2), 4-hydroxy-2, 6-dimethoxy-benzoic acid ( 3 ), 5-hydroxy-2-hydroxymethyl-γ-pyranone ( 4 ), 3, 5-dihydroxy-2-methyl-γ-pyranone(5),3,5-dihydroxy-γ-pyranone (6),daucosterol (7),4-hydroxybenzoic acid (8),pyrocatechol (9),β-sitosterol (10).Conclusion Compounds 1-7 have been obtained from this plant for the first time ,and 1-6 from the genus for the first time.The screening results of antitumor activity in vitro show that the half inhibitory concentration IC 50 of compound 2 for human osteosarcoma cells U20S and human neuroblastoma cells SY 5Y is 18.10 and 26.59 μg/ml respectively. Compound 5 can inhibit the growth of human breast cancer cells MCF-7 with a half inhibitory concentration IC 50 of 37.31μg/ml.

Effect of sorafenib combined with thalidomide on angiogenesis in chick chorioallantoic membrane / 临床肝胆病杂志

ObjectiveTo investigate the effect of sorafenib and/or thalidomide on angiogenesis in chick chorioallantoic membrane (CAM). MethodsWhite eggs incubated for 7 days were used to establish a CAM model. The model eggs were randomly divided blank control group, sorafenib group, thalidomide group, and sorafenib/thalidomide group. After treatment, each egg was incubated for another 2 days. The CAM samples were collected and fixed to take their pictures under a microscope, and the vascular coverage of each sample was calculated. Comparison between these groups was made by analysis of variance, and comparison between each two groups was made by SNK-q test. ResultsThe thalidomide group or sorafenib group had significantly lower vascular coverage than the blank control group (30.2%±2.9% or 26.5%±2.1% vs 38.3%±2.7%, P＜0.05). The sorafenib/thalidomide group had significantly lower vascular coverage than the thalidomide group or sorafenib group (12.6%±1.5% vs 30.2%±2.9% or 26.5%±2.1%, P＜0.05). ConclusionBoth sorafenib and thalidomide have a good anti-angiogenic effect on CAM, but a combination of the two drugs shows better efficacy.

Pharmacodynamic screening of anticancer agents used as c-Met inhibitor in vitro and in vivo / 天津医药

Objective To screen 8 series of LY compounds, c-Met tyrosine kinase inhibitors, and evaluate their anti-tumor effects in vitro and in vivo. Methods Preliminary screening was carried out by detecting the c-Met kinase phosphor?ylation inhibition activity of the compounds. CCK-8 assay was adopted for secondary anti-tumor screen of the selected com?pounds using MKN-45, U87MG, Caki-1 and PC-3 cell lines in vitro. The transplanted tumor model of U87MG cells in nude mice was established to evaluate the antitumor activity in vivo. Results Four compounds (LY22, LY25, LY28 and LY32) with better activities were selected by HTRF method, in which LY28 had better inhibitory effect on c-Met than that of Crizo?tinib. The above active compounds showed different degrees of inhibition on the four kinds of target cells (MKN-45, U87MG, Caki-1 and PC-3) detected by CCK-8 method, and the inhibitory effect of LY28 showed the most obvious. Antitumor activi?ty in vivo showed that LY28 can significantly inhibited tumor growth in a dose-dependent manner. The tumor inhibitory rate in high-dose of LY28 was 78.13%. Conclusion The compound LY28 has good antitumor activity in vitro and in vivo, which will be a new tyrosine kinase inhibitor.

Association between Chemotherapy-Response Assays and Subsets of Tumor-Infiltrating Lymphocytes in Gastric Cancer: A Pilot Study

PURPOSE: The purpose of this pilot study was to evaluate the association between adenosine triphosphate-based chemotherapy response assays (ATP-CRAs) and subsets of tumor infiltrating lymphocytes (TILs) in gastric cancer. MATERIALS AND METHODS: In total, 15 gastric cancer tissue samples were obtained from gastrectomies performed between February 2007 and January 2011. Chemotherapy response assays were performed on tumor cells from these samples using 11 chemotherapeutic agents, including etoposide, doxorubicin, epirubicin, mitomycin, 5-fluorouracil (5-FU), oxaliplatin, irinotecan, docetaxel, paclitaxel, methotrexate, and cisplatin. TILs in the tissue samples were evaluated using antibodies specific for CD3, CD4, CD8, Foxp3, and Granzyme B. RESULTS: The highest cancer cell death rates were induced by etoposide (44.8%), 5-FU (43.1%), and mitomycin (39.9%). Samples from 10 patients who were treated with 5-FU were divided into 5-FU-sensitive and -insensitive groups according to median cell death rate. No difference was observed in survival between the two groups (P=0.216). Only two patients were treated with a chemotherapeutic agent determined by an ATP-CRA and there was no significant difference in overall survival compared with that of patients treated with their physician's choice of chemotherapeutic agent (P=0.105). However, a high number of CD3 TILs was a favorable prognostic factor (P=0.008). Pearson's correlation analyses showed no association between cancer cell death rates in response to chemotherapeutic agents and subsets of TILs. CONCLUSIONS: Cancer cell death rates in response to specific chemotherapeutic agents were not significantly associated with the distribution of TIL subsets.

Evaluation of antimicrobial and antitumoral activity of Garcinia mangostana L. (mangosteen) grown in Southeast Brazil

PURPOSE: To characterize the anatomy of the fruit and leaf and the presence of phytocompounds. To evaluate the antitumor and antimicrobial activity of ethanolic extract of Garcinia mangostana L. (mangosteen) cultivated in southeastern Brazil. METHODS: Anatomical characterization and histochemical reactions were performed for structural identification and the presence of phytocompounds. Preparation of ethanolic extract of the fruit, leaf and resin of mangosteen. Culture B16-F10 melanoma cells for treatment with mangosteen ethanolic extract to determine cell viability by MTT and genotoxic effect by comet assay. Evaluation by antimicrobial activity against Staphylococcus aureus and Escherichia coli by agar diffusion test and by determination of Minimum Inhibitory Concentration (MIC). RESULTS: Our results showed many secretory canals in resin fruit and leaf; identifying lipids, starch, lignin and phenolic compounds. The leaf extract induced genotoxicity and apoptosis in B16-F10 cells, since the fragmentation of DNA in the comet assay. The ethanolic extract of mangosteen obtained in the resin, leaf and fruit showed antimicrobial activity against Staphylococcus aureus and Escherichia coli with a MIC at 0.1 mg/mL. CONCLUSION: In conclusion, we have demonstrated both antimicrobial and antitumor activity of ethanol extract of mangosteen emphasizing its therapeutic potential in infectious diseases and in cancer, such as melanoma. .

Evaluation of antitumoral and antimicrobial activity of Morinda lcitrifolia L. grown in Southeast Brazil

PURPOSE: To evaluate the antitumor and antimicrobial activity of ethanolic extract of Morinda citrifolia L. fruit cultivated in southeastern Brazil. METHODS: Preparation ethanolic extract of the fruit of Morinda citrifolia L. Culture of melanoma cells B16-F10 for treatment with ethanolic extract of Morinda citrifolia L. fruit to determine cell viability by MTT and determination temporal effect of ethanolic extract fruit on the cell growth B16-F10 for 8 days. Evaluation of antimicrobial activity of ethanolic extract fruit against Staphylococcus aureus and Escherichia coli by determination of Minimum Inhibitory Concentration (MIC). RESULTS: The ethanolic extract of Morinda citrifolia L. fruit (10mg/mL) decreased cellular activity and inhibited 45% the rate of cell proliferation of B16-F10 melanoma treated during period studied. The ethanolic extract of Morinda citrifolia L. fruit demonstrated antimicrobial activity inhibiting the growth of both microorganisms studied. Staphylococcus aureus was less resistant to ethanolic extract of Morinda citrifolia L. fruit than Escherichia coli, 1 mg/mL and 10 mg/mL, respectively. CONCLUSION: What these results indicate that the ethanolic extract of the fruit of Morinda citrifolia L. showed antitumor activity with inhibition of viability and growth of B16-F10 cells and also showed antibacterial activity as induced inhibition of growth of Staphylococcus aureus and Escherichia coli. .

Progress of methods for tumor chemosensitivity testing / 肿瘤研究与临床

Tumor chemosensitivity testing is very important for guiding individual chemotherapy.The more sensitive, faster and simpler methods for tumor chemosensitivity testing have been constantly developed.We reviewed the progress and status of methods for tumor chemosensitivity testing.

Combined oxaliplatin with adenosine triphosphate tumor chemosensitivity assay (ATP-TCA) for direction of indiyidual chemotherapy of hepatocellular carcinoma / 中国医师杂志

ObjectiveTo investigate the effectiveness of combined oxaliplatin regimen as adjuvant chemotherapy for hepatocellular carcinoma and to evaluate the efficacy of using adenosine triphosphate tumor chemosensitivity assay (ATP-TCA) for direction of individual chemotherapy.MethodsThe twenty-six patients with primary hepatocellular carcinoma were operated.Specimens were collected and adenosine triphosphate tumor chemosensitivity assay (ATP-TCA) was applied to evaluate the sensitiveness of chemotherapy agent(Adriamycin, Mitomycin, Mitoxantrone, Oxaliplatin, Irinotecan, 5-FU, Gemzar, Carboplatin, Cisplatin, Docetaxel and Etoposide).Sensitive group (SG) was from from 11 patients who were sensitive to oxaliplatin, and control group was from the other 16 patients who were not sensitive to oxaliplatin.All the twenty-six patients received oxaliplatin combined with 5-FU or capecitabine regimen chemotherapy.The effectiveness (CR,PR,SD,PD,ORR,OS and DFS) of the regimen according to RECIST criteria and WHO criteria for anticancer drugs toxicity and efficacy of ATP-TCA were evaluated.ResultsTwenty-six patients were successfully evaluated.In SG, six patients obtained complete remission(CR), three got partial remission(PR), one got stable disease (SD) and one patient got progression disease (PD).While in control group,four patients obtained CR,two patients got PR, five patients got SD and four got PD.No significant differences were found in overall survival (OS, P = 0.1116) and disease-free survival (DFS, P = 0.2328)between sensitive group and control group.But significant differences were found in overall response rate (ORR) (81.8％ vs 40.0％, P =0.0401) between two groups.Common toxicities were as follows:I to Ⅱdegree of myelosuppression was 53.8％, I to Ⅱ degree of gastrointestinal tract response was 50％, I to Ⅱ degree of liver function damage was 57.7％ and I to Ⅱ degree of neuropathy was 23.1％, respectively.Most of these toxicities were tolerable at grade 1 ～ 2.No significant differences were found in the toxicities between two groups.ConclusionsCombined oxaliplatin regimen might be an effective choice for adjuvant chemotherapy for HCC, which has with tolerable systemic toxicity.Application of ATP-TCA system might further improve the efficacy of this regimen by selecting right candidate.

Predicting clinical chemo-sensitivity of primary ovarian cancer using adenosine triphosphate-tumor chemosensitivity assay combined with detection of drug resistance genes / 中华妇产科杂志

Objective To predict clinical chemotherapy sensitivity of primary ovarian cancer by jointing adenosine triphosphate(ATP) - tumor chemo-sensitivity assay(TCA) method in vitro and detection of drug resistance genes, provide reference for clinical treatment. Methods Forty-seven primary epithelial ovarian tumor samples were collected from the patients who received cytoreductive surgery. Viable ovarian cancer cells obtained from malignant tissue were tested for their sensitivity to carboplatin (CBP), cisplatin (DDP), paclitaxel(PTX) and CBP + PTX using ATP-TCA method in vitro; at same time, real-time quantitative PCR was used to analysis BRCA1 and ERCC1 mRNA relative expression in forty-six specimens (1 frozen tumor samples mRNA were not detected due to serious degradation). The relationship between ATP-TCA test results, clinical indicators, and the effectiveness of the joint prediction on clinical chemosensitivity by combining these two methods were statistically analyzed using chi-square test. Results (1)The results showns that three programs of DDP,CBP and PTX + CBP were significantly related with clinical results(P<0.05) in vitro, in which the compliance rate in PTX + CBP program was the highest 83%(39/47) ,and the predictive sensitivity, predictive specificity, positive predictive value, negative predictive value and predictive accurate rate were 90%,71%,84% and 80% ,respectively.PTX + CBP combined in vitro test results was also related with residual tumor size and neoadjuvant chemotherapy, which was more prone to drug resistance with residual tumor larger than 2 cm (P = 0. 023) and with neoadjuvant chemotherapy (P = 0.011). (2) BRCA1 mRNA expression levels in the clinical-resistant group and the clinical-sensitive group was 0.673 ± 2.143 and - 1.436 ± 2.594 (P=0.008), ERCC1 mRNA expression levels in the clinical-resistant group and the clinical-sensitive group was -0.529 ± 1.982 and - 3.188 ±2.601 (P =0.001). There were also significant correlation among the expression levels of BRCA1 ,ERCC1 mRNA and clinical efficacy (P<0.01). (3)ATP-TCA and detection of drug resistance genes combined to predict the clinical application of PTX + CBP resistance may occur in 8/9 cases. Conclusions ATP-TCA may be an ideal method of in vitro drug sensitivity testing method, which could effectively predict clinical chemotherapy sensitivity. Combination of the drug-resistant associated genes detection method and the ATP-TCA method can increase the predictive effectiveness of ovarian cancer chemosensitivity and guide individual chemotherapy of ovarian cancer.

Isolation and culture of cancer stem cells from ovarian cancer tissues and malignant ascites / 肿瘤

Objective: To isolate and culture tumor stem cells from ovarian cancer tissues and malignant ascites, and to analyze the cellular phenotype and drug sensitivity. Methods: The primary tumor tissues and the malignant ascites samples from one patient with ovarian adenocarcinoma were collected, and the formations of float-growing spheres was developed with serum-free medium culture. The expressions of stem cell markers were evaluated by flow cytometry (FCM) and immunofluorescent staining. The sensibilities of sphere cells and differentiated cells to paclitaxol and carboplatin were measured by in vitro drug sensitivity assay. Results: The ovarian cancer cells in both tumor tissues and malignant ascites displayed an ability to generate spheres cultured in serum-free medium. These sphere-forming cells exhibited the phenotype of CD44+CD24- and also expressed stem cell markers including Oct4, Nanog and Sox2. These sphere cells from tumor tissues and malignant ascites became differentiated after culture in serum-containing medium for 3 weeks, and the fractions of CD44+ and CD24- cells reduced by 47% and 3%, respectively. The differentiated cells showed increased sensitivities to paclitaxol and carboplatin as compared with the sphere cells (P<0.01). Conclusion: The ovarian cancer stem cells exhibit the phenotype of CD44+CD24-, which is characterized by expressions of several stem cell markers including Oct4 and a high level of drug resistance. Copyright© 2011 by TUMOR.

Clinical research on chemotherapy of recurrent and refractory non-Hodgkin lymphoma directed by ATP bioluminescence chemosensitivity assay in vitro / 白血病·淋巴瘤

ObjectiveTo investigate the clinical value of ATP bioluminescence tumor chemosensitivity assay (ATP-TCA) for recurrent and refractory non-Hodgkin lymphoma (NHL) specimens in vitro.Methods Thirty-four freshlytaken recurrent andrefractoryNHL specimens weretestedin vitro for cancer chemosensitivity by ATP-TCA.ResultsDrug sensitivity of NHL specimens had heterogeneity.Different drugs had different tumor growth inhibition ratio in vitro.Response rate (RR) of the patients receiving chemotherapy according to in vitro assay was 82.4 ％ (28/34),complete response rate (CR) was 52.9 ％ (18/34).In DICE group RR was 60.0 ％ (18/30),CR rate was 33.3 ％ (10/30).In GDP group RR was 62.3 ％ (33/53),CR rate was 26.4 ％ (14/53).In ATP-TCA group RR was significantly higher than those in DICE and GDP groups (x2 =3.93,P =0.047; x2 =3.98,P =0.046).ConclusionThe results of ATP-TCA assay are correlated well with clinical treatment responses.The assay may be an important and useful method for individual-based chemotherapy of cancers.

Predictive effect of combined evaluation of XRCC1, XPD and GSTP1 single nucleotide polymorphisms in platinum based chemotherapy / 中国医师杂志

Objective To investigate the predictive value of combined analysis on single nucleotide polymorphisms (SNPs) of X-ray cross-complementing1 ( XRCC1 ) gene 194 and 399 codon,xeroderma pigmentosum group D (XPD) gene 312 codon and glutathione S-transferase P1 (GSTP1) gene 105 codon in platinum based chemotherapy.Methods Direct sequencing was performed to detect XRCC1,XPD and GSTP1 genotypes in peripheral blood from 50 cancer patients receiving platinum-based chemotherapy.Genetic polymorphisms of these genes related to sensitivity of platinum were reviewed.Results Favorable genotypes were Arg/Trp and Trp/Trp in XRCC1 194 codon,Arg/Arg in XRCC1 399 codon,Asn/Asn in XPD 312 codon and Val/Val in GSTP1 105 codon.The response rate to chemotherapy was 57.1％,75.0％,60.9％,85.7％ and 87.5％,respectively.The response rate for patients possessing ≥2 favorable genotypes and those possessing 1 or 0 favorable genotype was 78.9％,36.4％ and 0,respectively.Patients possessing ≥2 favorable genotypes demonstrated higher sensitivity to platinum based chemotherapy,compared with those possessing 1 or 0 favorable genotype ( x2 =25.79,P ＜ 0.01 ).Conclusions Combination analysis of genomic polymorphisms of XRCC1,XPD and GSTP1 may be useful in predicting sensitivity of platinum based chemotherapy.

Effect of ginsenoside Rg3 on Cx26 gene expression and gap junctional intercellular communication in human breast cancer cell line MCF-7 / 肿瘤研究与临床

Objective To study the function of ginsenoside Rg3 on proliferation in human breast cancer cell line MCF-7 and effect of ginsenoside Rg3 on Cx26 gene expression and gap junctional intercellular communication (GJIC) in MCF-7, cultured in vitro. MethodsHuman breast cancer cells MCF-7 was exposed to ginsenoside Rg3 at differential concentrations for 24 h, respectively. The cell proliferation inhibition was measured by 3-[4,5-dimethylthiazo-2-yl]-2,5 diphenyl tetrazolium bromide (MTT) assay. The expression of Cx26 mRNA was measured by RT-PCR in experimental groups and control goup. The GJIC function of MCF-7 cell was examined with scrape-loading dye transfer assay.ResultsHuman breast cancer cell line MCF-7 was exposed to ginsenoside Rg3 at a concentration of 10, 20, 40, 80, 160 μg/ml, respectively.The inhibition ratio was 3.1%, 5.2 %, 16.0 %, 26.3 %, 29.1% respectively after 24 h. Compared with control group, the concentration of 40 μg/ml above could significantly inhibit MCF-7 cell proliferation (P ＜0.05), so the experimental groups were exposed to ginsenoside Rg3 at a concentration of 40, 80, 160 μg/ml,respectively. The expression of Cx26 mRNA in every experimental group compared with control group was enhanced when MCF-7 cell was exposed to ginsenoside Rg3 at a higher concentration. It was observed that Lucifer yellow fluorescent staining was limited to a single cell in control group through fluorescent microscope,but Lucifer yellow fluorescent transfered through gap junction cells to neighboring cells, then came into being flake fluorescent staining in experiment groups. ConclusionGinsenoside Rg3 can enhance the expression of Cx26 mRNA in MCF-7 cell and restore the gap junctional intercellular communication, which may be one of important mechanisms of ginsenoside Rg3 in antitumor.

ERCC1 and BRCA1 mRNAs expression levels in malignant pleural and peritoneal effusions are associated with chemosensitivity to cisplatin in vitro / 肿瘤

Objective:The aim of this study was to investigate the association of mRNA expressions of ERCC1 (excision repair cross-complementing group 1) and BRCA1 (breast cancer 1) with chemosensitivity to cisplatin in malignant pleural and peritoneal effusions.Methods:Malignant pleural and peritoneal effusions were collected from 46 patients diagnosed with stage Ⅳ malignant tumor, prospectively. The tumor cells were isolated and the sensitivity of tumor cells to cisplatin was detected by adenosine triphosphate-bioluminescence assay (ATP-TCA). Real-time quantitative PCR was used to determine the mRNA expressions of ERCC1 and BRCA1. Results:The expression level of ERCC1 mRNA was negatively correlated with sensitivity of non-small cell lung cancer (NSCLC) to cisplatin (P= 0.001, r=0.685). BRCA1 mRNA expression level had negative correlation with sensitivity to cisplatin in both NSCLC (P=0.014, r=0.541) and gastric cancer (P=0.002, r=0.625). A significant interaction was found between the effects of ERCC1 and BRCA1 mRNA expressions on sensitivity to cisplatin (P=0.010 for all patients;P=0.027 for gastric cancer patients).Conclusion:ERCC1 and BRCA1 mRNA expression levels correlated with ex vivo chemosensitivity of tumor cells to cisplatin in malignant pleural and peritoneal effusions. Detection of both ERCC1 and BRCA1 may have a higher reliability in predicting the sensitivity of tumor cells to cisplatin than detection of single ERCC1 or BRCA1 expression.

The drug sensitivity of human lung adenocarcinoma stem cells to cisplatin and carboplatin / 肿瘤

Objective:To analyze the drug sensitivity of human lung adenocarcinoma stem cells (LASC) to cisplatin (DDP) and carboplatin (CBP). Methods:Human lung adenocarcinomaic cells SPC-A1,AG,and CPA-Y2 were treated with DDP and CBP. The cell viability of cells was detected by CCK-8 assay. The phenotypic characteristics of drug surviving cells(DSCs)were determined by immunofluorescence staining. The LASC population was then separated by magnetic-activated cell sorting method. The LASC in DSCs was traced by using green fluorescence protein (GFP). The drug sensitivity of DSCs to DDP and CBP was analyzed.Results:The LASC exhibited the phenotypes of bronchioalveolar stem cells (BASC, OCT4~+CCSP~+SP-C~+). After mixture of CD221~+LASC with CD221~-lung adenocarcinoma differentiated cells, the DSC population showed OCT4~+BASC phenotypes. These DSCs were significantly resistant to DDP and CBP.Conclusion:LASC has a high resistance to DDP and CBP. This may be the reason for tumor recurrence after chemotherapy.

Associação do quimioterápico daunorrubicina a uma nanoemulsão rica em colesterol: estudos de regressão tumoral e farmacocinética / Association of the chemotherapeutic agent daunorubicin a cholesterol-rich nanoemulsion: regression studies pharmacokinetics and tumor

A nanoemulsão lipídica (LDE) se concentra nas células neoplásicas e pode ser utilizada como transportador de derivado lipofílico da daunorrubicina, como o Noleil- daunorrubicina (oDNR). Neste estudo, a LDE-oDNR foi preparada por homogeneização em alta pressão e sua toxicidade e atividade anti-tumoral testadas. A associação LDE-oDNR teve rendimento elevado e permaneceu estável por longo período. Em camundongosC57BL/6J, a dose máxima tolerada (DMT) foi 65 vezes maior e a DL50 48 vezes maior no tratamento LDE-oDNR comparado ao tratamento DNR comercial, resultando em alta redução da toxicidade. Em camundongos implantados com células de melanoma B16, a preparação LDE-oDNR (7,5 mol/kg) levou a redução de 59 ± 2% do crescimento do tumor comparado a redução de 23 ± 2% para o tratamento DNR comercial na mesma dose (p<0,001). A probabilidade de sobrevida teve aumento pronunciado nos animais tratados com LDE-oDNR comparado à DNR comercial (p <0,01). Além disso, apenas 30% dos animais portadores de melanoma submetidos ao tratamento com LDE-oDNR apresentaram metástases, comparado a 82% quando tratados com DNR comercial. Uma forte redução de toxicidade também foi observada pela redução da anemia e leucopenia nos animais tratados com LDE-oDNR, em comparação com DNR comercial. A preparacao LDE-oDNR foi eficaz também no quadro de trombocitose induzida por tumor. Os testes com fragmentos extraídos de tumores dos animais tratados mostraram que a LDE-oDNR foi mais eficaz na destruição das células neoplásicas comparado ao tratamento DNR comercial (9% de células viáveis com tratamento LDE-oDNR, 27% sob tratamento DNR). O estudo mostrou que o tratamento proposto com o derivado ODNR associado à nanoemulsão (LDE-oDNR) é efetivo no combate às células tumorais, seletivo, menos tóxico e melhor tolerado. Os estudos de farmacocinética e biodistribuição somam a este protocolo informações importantes relacionadas às propriedades de absorção, distribuição, metabolismo e excreção...

A lipidic nanoemulsion (LDE) that concentrates in neoplastic cells can be used as vehicle to daunorubicin lipophylic derivatives, such as N-oleyl-daunorubicin (oDNR). Here, LDE-oDNR was prepared by high pressure homogenization to test toxicity and anti-tumor activity. LDE-oDNR association yield was high and stable for long period. In mice, maximum tolerated dose was 65 and LD50 was 48-fold greater in LDE-oDNR than in commercial DNR treatment, showing very strong toxicity reduction. In melanoma B16-tumor bearing mice, LDE-oDNR (7.5 mol/Kg) reduced tumorgrowth by of 59±2%, and DNR by only 23±2% at same dose level (p<0.001). Survival was pronouncedly increased in LDE-oDNR compared to DNR treatment (p<0.01). Furthermore, the number of melanoma-bearing mice with metastasis was 30% under LDE-oDNR, compared to 82% under DNR treatment. Strong reduction of toxicity was also observed by reduction of anemia and leucopenia under LDE-oDNR, compared to commercial DNR tumor-induced thrombocytosis was more effective with LDE-oDNR than with DNR. Tests with fragments extracted from tumors of treated animals showed that LDE-oDNR was more effective in killing neoplastic cells than DNR (9% of viable cells under LDE-oDNR; 27% under DNR). The pharmacokinetics and biodistribution studies add important information to this protocol related to the properties of absorption, distribution, metabolism and excretion of the formulation under study compared to free DNR. The remarkable toxicity reduction and increase in pharmacological action supports novel LDE-oDNR as a promising weapon in cancer treatment.