Efficacy and safety of second-line camrelizumab combined with apatinib and chemotherapy in patients with advanced lung adenocarcinoma: A prospective, open-label, multicentric study.

OBJECTIVES: Camrelizumab (a programmed cell death protein 1 inhibitor) and apatinib (an angiogenesis inhibitor) are considered as potential treatments for advanced lung adenocarcinoma (LUAD). This study aimed to evaluate the efficacy and safety of second-line camrelizumab combined with apatinib and chemotherapy (albumin-bound paclitaxel, docetaxel, or pemetrexed) in patients with advanced LUAD. METHODS: Twenty-nine patients with advanced LUAD underwent second-line camrelizumab combined with apatinib and chemotherapy were enrolled in this prospective, open-label, multicentric study. Follow-up with a median duration of 18.0 months was conducted. RESULTS: There were 0 (0.0 %), 11 (37.9 %), 14 (48.4 %), and 3 (10.3 %) patients achieving complete response, partial response, stable disease, and progressive disease, respectively. Meanwhile, treatment response was not evaluated in 1 (3.4 %) patient. The objective response and disease control rates were 37.9 % and 86.3 %, respectively. In terms of survival, the median (95 % confidence interval) progression-free survival (PFS) was 11.1 (5.2-17.0) months, with 1-year and 2-year PFS rates of 40.4 % and 20.5 %, respectively. The median overall survival (OS) was not reached; the 1-year and 2-year OS rates were 72.0 % and 64.8 %, respectively. Current treatment cycles ≥ 8 were associated with better PFS and OS (both P < 0.001). In addition, 21 (72.4 %) patients experienced at least one treatment-emergent adverse event (TEAE), which was mostly of grade I and II. The most commonly occurring TEAE was leukopenia (17.2 %), liver dysfunction (17.2 %), hypothyroidism (13.8 %), hand-foot syndrome (13.8 %), and thrombocytopenia (13.8 %). CONCLUSION: Second-line camrelizumab combined apatinib and chemotherapy might serve as a potential treatment with acceptable safety in patients with advanced LUAD.

KIAA1199 promotes oxaliplatin resistance and epithelial mesenchymal transition of colorectal cancer via protein O-GlcNAcylation.

Oxaliplatin is a commonly used platinum drug for colorectal cancer (CRC). However, the treatment of CRC by oxaliplatin usually fails because of drug resistance, which results in a huge challenge in the therapy of CRC. Elucidation of molecular mechanisms may help to overcome oxaliplatin resistance of CRC. In our study, we revealed that KIAA1199 can promote oxaliplatin resistance of CRC. Mechanistically, KIAA1199 prevents oxaliplatin mediated apoptosis via up-regulated PARP1 derived from reduced endoplasmic reticulum stress induced by protein O-GlcNAcylation. In the meantime, KIAA1199 can also trigger epithelial mesenchymal transition by stabilizing SNAI1 protein via O-GlcNAcylation. Therefore, KIAA1199 has great potential to be a novel biomarker, therapeutic target for oxaliplatin resistance and metastasis of CRC.

Pretreatment BAN Score Based on Body-mass-index, Albumin and Neutrophil-lymphocyte Ratio Could Predict Long-term Survival for Patients with Operable Esophageal Squamous Cell Carcinoma.

Background: The present study was designed to examine the prognostic value of a systemic inflammation marker-BAN score, which was established based on body-mass-index (BMI), albumin (ALB) and neutrophil-lymphocyte ratio (NLR) in resectable esophageal squamous cell carcinoma (ESCC) patients. Methods: A total of 420 newly diagnosed ESCC patients in our hospital between January 2008 and December 2013 were included. Their baseline characteristics were retrospectively reviewed and collected. BAN score was calculated as (BMI × ALB/ NLR). The optimal cutoff value for BAN score was defined as 28.0 in terms of survival. Patients were then allocated to high BAN (≥ 28.0) and low BAN (< 28.0) score groups. Results: Pretreatment BAN score was significantly associated with tumor length, white blood cell count, BMI, ALB and NLR levels. However, no significant difference was observed in patients' age, gender, tumor location, degree of differentiation, depth of invasion, lymph node involvement, tumor-node-metastasis (TNM) stage or other variables between groups. Moreover, those with high pretreatment BAN scores (≥ 28.0) tended to have favorable disease free survival (DFS) [hazard ratio (HR), 0.650; 95% confidence interval (CI), 0.481-0.877; P = 0.005] and overall survival (OS) (HR, 0.608; 95% CI, 0.445-0.829; P = 0.002) by univariate analysis. Furthermore, multivariate Cox regression analysis suggested that high BAN score (≥ 28.0) could serve as an independent predictor for both DFS (HR, 0.726; 95% CI, 0.532-0.993; P = 0.045) and OS (HR, 0.670; 95% CI, 0.485-0.927; P = 0.016). Conclusions: Pretreatment BAN score could independently predict long-term survival for resectable ESCC patients.

Large Scale, Multicenter, Prospective Study of Apatinib in Advanced Gastric Cancer: A Real-World Study from China.

BACKGROUND: In China, gastric cancer (GC) ranks second in incidence and mortality. Over 80% of patients with GC were diagnosed at an advanced stage with poor clinical outcome. Chemotherapy was the mainstream treatment with limited benefit. Apatinib, an inhibitor of targeting vascular endothelial growth factor receptor 2 (VEGFR2), has been approved for third-line treatment of advanced gastric cancer. However, the data of apatinib treatment in the real-world setting are limited. In this real-world study, we aimed to understand the current treatment pattern of apatinib, investigate the effectiveness and safety of apatinib in real-world settings, and explore the potential factors associated with the clinical outcomes. METHODS: This was a prospective, multicenter observational study in a real-world setting. Patients aged ≥18 years with histologic diagnosis of advanced GC were eligible for enrollment. The eligible patients received either apatinib monotherapy or apatinib plus chemotherapy by physician's discretion. Apatinib treatment could be used as first-line, second-line, or third-line and above therapy. The primary endpoint was progression-free survival (PFS). The secondary endpoints were overall survival (OS), ORR, DCR, and safety profile. RESULTS: A total of 737 patients with advanced gastric cancer treated with apatinib were included in the FAS population. A total of 54.9% patients used apatinib monotherapy and 45.1% patients used apatinib combination therapy. A total of 44.1% patients received apatinib in first-line treatment, 28.2% in second-line, and 27.7% in third-line and above. In first-line treatment, the objective response rate (ORR) was 9.09% and 16.42% in apatinib monotherapy and combination therapy groups, and disease control rate (DCR) was 78.41% and 89.29%, respectively. Patients who received combination therapy achieved significantly longer median progression-free survival (mPFS; 6.18 vs 3.52 months, p<0.01) and median overall survival (mOS; 8.72 vs 5.92 months, p<0.01) compared with monotherapy. In second-line and third-line therapy, combination therapy showed a better trend in tumor response and survival outcomes compared with monotherapy. For all patients, apatinib combined with paclitaxel were associated with longer mPFS compared with other combinations (8.88 vs 6.62 months). Multivariate analysis showed that combination with paclitaxel (p=0.02) and experience of apatinib-related specific AEs (p<0.01) were independent predictors for PFS and OS. The safety profile was tolerable and no unexpected adverse events were reported. CONCLUSION: In a real-world setting, apatinib showed a favorable effectiveness and safety profile in patients with advanced gastric cancer. Apatinib combination therapy, especially combined with paclitaxel, might lead to better survival benefit in first-line treatment. Combination with paclitaxel and the occurrence of apatinib-specific AEs were independent factors associated with better survival outcomes. TRIAL REGISTRATION: NCT03333967.

Survival benefits of palliative gastrectomy in stage IV gastric cancer: a propensity score matched analysis.

BACKGROUND: This study aimed to investigate the effect of palliative gastrectomy on survival in stage IV gastric cancer. METHODS: Patients diagnosed with stage IV gastric cancer between 2010 and 2016 were selected from the Surveillance, Epidemiology, and End Results (SEER) database. Survival curves were estimated by the Kaplan-Meier method before and after propensity score matching (PSM). Univariate and multivariate Cox analyses were performed to evaluate risk factors for survival in patients who underwent palliative gastrectomy. RESULTS: We examined 6,529 patients with stage IV gastric cancer, of which 625 underwent palliative gastrectomy. Using a 1:2 PSM, the 625 patients were matched with 1,250 patients from the no gastrectomy group. The overall survival was higher in the gastrectomy group before [hazard ratio (HR) =0.57, 95% confidence interval (CI): 0.53-0.62, P<0.0001] and after PSM (HR =0.51, 95% CI: 0.46-0.57, P<0.0001). Multivariate Cox analysis confirmed the survival benefits of palliative gastrectomy and chemotherapy. Older age, over-lapping lesions, non-adenocarcinomas, higher tumor grade, and lung metastasis significantly increased the risk of mortality. In the gastrectomy group, patients aged ≥80 years, diagnosed with grades 3/4 non-adenocarcinomas, or with lung metastasis showed poorer prognosis. However, chemotherapy could improve the survival of these patients. CONCLUSIONS: Palliative gastrectomy provides survival benefits to stage IV gastric cancer patients. However, age, tumor grade, tumor histology, and lung metastasis status should be considered while making a decision regarding gastrectomy. Chemotherapy should also be recommended for these patients.

Surgery on primary tumor shows survival benefit in selected stage IV colon cancer patients: A real-world study based on SEER database.

Objectives: Most patients with stage IV colon cancer did not have the opportunity for curative surgery, only selected patients could benefit from surgery. This study aimed to determine whether surgery on the primary tumor (SPT) should be performed in patients with stage IV colon cancer and how to select patients for SPT. Methods: This study included 48,933 patients with stage IV colon cancer who were identified in the Surveillance, Epidemiology and End Results (SEER) database between 1998 and 2015. Propensity score matching (PSM) analysis was adopted to balance baseline differences between SPT and non-surgery groups. Kaplan-Meier (K-M) curves were utilized to compare the overall survival (OS). Prognostic nomograms were generated to predict survival based on pre- and post-operative risk factors. Patients were divided into low, middle, and high mortality risk subsets for OS by X-tile analyses based on scores derived from above nomograms. Results: Patients with SPT had a significantly longer OS than those without surgery, regardless of the metastatic sites and diagnostic years. Nomograms, according to the pre- and post-operative risk factors, showed moderate discrimination (all C-indexes above 0.7). Based on X-tile analyses, low mortality risk subset (post-operative score ≤ 22.3, preoperative score ≤ 9.7) recommended for SPT, and high mortality risk was not. Conclusions: SPT led to prolonged survival in stage IV colon cancer. Our nomograms would help to select suitable patients for SPT.

Survival value of primary tumour resection for stage IV non-small-cell lung cancer: A population-based study of 6466 patients.

INTRODUCTION: For stage IV non-small-cell lung cancer (NSCLC) patients, surgical resection of primary tumour was rarely recommended. OBJECTIVES: We conducted this population-based study to demonstrate the survival value of primary tumour resection (PTR) for stage IV (NSCLC). METHODS: The Surveillance, Epidemiology and End Results (SEER) database was searched for selecting stage IV NSCLC patients. The patients were matched according to age, gender, grade, primary tumour site, histopathological type, tumour size and regional lymph nodes metastasis by propensity score matching (PSM) analysis. Kaplan-Meier curves were presented to show the survival differences between resection group and non-resection group. Risk factors which were supposed to influence survival outcome were investigated using a Cox proportional hazard regression model. And a nomogram was performed to present prognostic factors for stage IV NSCLC patients. RESULTS: 6466 patients diagnosed from 2004 to 2015 were included in survival analyses after PSM. The median overall survival (OS) for overall patients with resection was 27 months, much longer than those without resection (8 months). And this trend remained in subgroup analyses, including different histopathological types and distant metastases (All P values < 0.001). Younger age, race other than white and black, female, grade 1/2 (G1/G2), PTR, chemotherapy, no other distant metastases, smaller tumour size and no regional lymph node metastases were favourable prognostic factors for stage IV NSCLC. A predictive nomogram was conducted based on above risk factors. CONCLUSION: PTR prolonged survival of stage IV NSCLC patients. And PTR should be considered in clinical practice for stage IV NSCLC.

MicroRNA-215-3p Suppresses the Growth, Migration, and Invasion of Colorectal Cancer by Targeting FOXM1.

Previous investigations have indicated that microRNA-215-3p is dysregulated in many kinds of cancers and functions as oncogene or tumor suppressor. However, the potential role of microRNA-215-3p in the progression of colorectal cancer remains not well known. Herein, we demonstrated that microRNA-215-3p was downregulated in human colorectal cancer tissues and was reversely correlated to the lymph node metastasis of colorectal cancer. Overexpression of microRNA-215-3p inhibited the clonogenic abilities and metastasis-relevant traits of colorectal cancer cell in vitro. Consistently, upregulation of microRNA-215-3p inhibited the growth and metastasis of colorectal cancer cell in vivo. Forkhead box protein M1 was identified as a direct target of microRNA-215-3p and reexpression of forkhead box protein M1 reversed the suppressive impacts of microRNA-215-3p on the growth, mobility, and invasion abilities of colorectal cancer cell. Altogether, these results revealed the vital role of microRNA-215-3p in the tumorigenesis and metastasis of colorectal cancer.

Nomograms for predicting survival in patients with metastatic gastric adenocarcinoma who undergo palliative gastrectomy.

BACKGROUND: Recently, evidence has emerged that palliative gastrectomy in patients with stage IV gastric cancer may offer some survival benefits. However, the decision whether to perform primary tumor surgery remains challenging for surgeons, and investigations into models that are predictive of prognosis are scarce. Current study aimed to develop and validate prognostic nomograms for patients with metastatic gastric adenocarcinoma treated with palliative gastrectomy. METHODS: The development dataset comprised 1186 patients from the Surveillance, Epidemiology, and End Results Program who were diagnosed with metastatic gastric adenocarcinoma in 2004-2011, while the validation dataset included 407 patients diagnosed in 2012-2015. Variables were incorporated into a Cox proportional hazards model to identify independent risk factors for survival. Both pre- and postoperative nomograms for predicting 1- or 2-year survival probabilities were constructed using the development dataset. The concordance index (c-index) and calibration curves were plotted to determine the accuracy of the nomogram models. Finally, the cut-off value of the calculated total scores based on preoperative nomograms was set and validated by comparing survival with contemporary cases without primary tumor surgery. RESULTS: Age, tumor size, location, grade, T stage, N stage, metastatic site, scope of gastrectomy, number of examined lymph node(s), chemotherapy and radiotherapy were risk factors of survival and were included as variables in the postoperative nomogram; the c-indices of the development and validation datasets were 0.701 (95% confidence interval [CI]: 0.693-0.710) and 0.699 (95% CI: 0.682-0.716), respectively. The preoperative nomogram incorporated age, tumor size, location, grade, depth of invasion, regional lymph node(s) status, and metastatic site. The c-indices for the internal (bootstrap) and external validation sets were 0.629 (95% CI: 0.620-0.639) and 0.607 (95% CI: 0.588-0.626), respectively. Based on the preoperative nomogram, patients with preoperative total score > 28 showed no survival benefit with gastrectomy compared to no primary tumor surgery. CONCLUSIONS: Our survival nomograms for patients with metastatic gastric adenocarcinoma undergoing palliative gastrectomy can assist surgeons in treatment decision-making and prognostication.

Prognostic value of DNA aneuploidy in gastric cancer: a meta-analysis of 3449 cases.

BACKGROUND: DNA aneuploidy has attracted growing interest in clinical practice. Nevertheless, its prognostic value in gastric cancer patients remains controversial. This meta-analysis aims to explore the impact of DNA ploidy status on the survival of gastric cancer patients. METHODS: We used PubMed and Web of Science databases to retrieve relevant articles. The correlation between DNA aneuploidy and the clinicopathological features of gastric cancer, such as stage, depth of invasion (T), lymph node metastasis (N), distant metastasis (M), differentiation (G), tumor types (Lauren classification) and overall survival (OS) were evaluated. Hazard ratios (HRs) with corresponding 95% confidence intervals (CIs) were collected carefully from each article OS was presented with HRs. The relationships between DNA aneuploidy and each characteristic were analyzed using risk ratios (RR) and a 95% confidence interval (CI). Significance was established using P < 0.05. Funnel plot was conducted to detect the publication bias. RESULTS: After careful selection, 25 studies involving 3449 cases were eligible for further analyses. Patients with DNA aneuploidy were considered at risk of more advanced stages (stage III-IV vs. stages I-II, RR = 1.23; 95% CI, 1.07 to 1.42; P = 0.003), lymph node metastasis (N+ vs. N-: RR = 1.43; 95% CI, 1.12 to 1.82, P = 0.004), and intestinal tumor type (intestinal vs. diffuse: RR = 1.45; 95% CI, 1.02 to 2.06; P = 0.04). And an adverse relation was observed between DNA aneuploidy and tumor differentiation. While no association was found between DNA aneuploidy and distant metastasis (P = 0.42) nor depth of tumor invasion (P = 0.86). Regarding overall survival, aneuploid tumors were associated with worse survival in all patients (P < 0.00001). CONCLUSIONS: We found that DNA aneuploidy was an important predictor for gastric cancer patients, and should be used as a potential biomarker for further classification in gastric cancer.

Prognostic value of Ki-67 in stage I non-small-cell lung cancer: A meta-analysis involving 1931 patients.

There is growing interest in exploring the prognostic value of Ki-67 in non-small-cell lung cancer (NSCLC). However, whether Ki-67 can be regarded as a routine biomarker in clinical practice is still under debate. The present meta-analysis investigated the relationship between Ki-67 and the overall survival (OS) or disease-free survival (DFS) of patients suffering from stage I NSCLC. We searched the Web of Science, Cochrane, and PubMed databases to extract eligible articles. In total, 15 studies involving 1931 patients were included. Pooled hazard ratio (HR) analysis revealed that patients with high Ki-67 labeling index (LI) had poorer OS (HR = 1.95, 95% confidence interval (CI) = 1.43-2.66, P < 0.0001) and DFS (HR = 3.12, 95% CI = 2.17-4.48, P < 0.00001) than those with low Ki-67 LI. In subgroup analysis, high Ki-67 LI was significantly associated with poor prognoses in stage I adenocarcinoma. In future studies, a consensus for the optimal cutoff value for high Ki-67 LI needs to be explored and demonstrated in stage I NSCLC patients.

Expression and clinical significance of Vimentin and E-cadherin in breast cancer tissues / 中国肿瘤生物治疗杂志

@#Objective: To study the expression and clinical significance of Vimentin and E-cadherin in human breast cancer tissues. Methods: : The clinical data of 56 cases of breast cancer patients, who underwent radical mastectomy in Chaohu Hospital Affiliated to Anhui Medical University from January 2014 to January 2016, were retrospectively analyzed. The protein and mRNAexpressions of Vimentin and E-cadherin in breast cancer tissues were detected by immunohistochemistry and qPCR, respectively; and the relationship between the expression of Vimentin and E-cadherin in breast cancer tissues and the clinicopathological characteristics was analyzed. Logistic multivariate regression was used to analyze the independent factors affecting the protein expressions of Vimentin and E-cadherin. Spearman was used to analyze the correlation between Vimentin and E-cadherin. Kaplan-Meier was used to analyze the relationship between protein expressions of Vimentin, E-cadherin and prognosis. ROC curve was used to analyze the diagnostic value of Vimentin and E-cadherin on prognosis. Results: The rates of breast cancer tissues with high positive expression of Vimentin and E-cadherin were 76.79% and 19.64%, respectively.Among them, 47 cases (47/56, 83.93%) of breast cancer tissues showed significantly higher Vimentin mRNA expression than adjacent tissues (P<0.05), and 46 cases (46/56, 82.14%) of breast cancer tissues showed significantly lower Ecadherin mRNA expression than adjacent tissues (P<0.05). Vimentin protein expression was associated with tumor size, lymph node metastasis, vascular invasion, histological grade, clinical stage, molecular typing, Ki67+, ER-, PR- and HER2- expression (P<0.05). And E-cadherin protein expression was associated with lymph node metastasis, vascular invasion, histological grade, clinical stage, molecular typing, Ki67+, ER-, PR- and HER2- expression (P<0.05). Tumor size, lymph node metastasis, vascular invasion, histological grading, clinical staging, molecular typing, Ki67+, ER-, PR- and HER2- expression were all independent factors affecting the expression of Vimentin and E-cadherin (P<0.05). There was a negative correlation between Vimentin and E-cadherin expression (P<0.05). The 3-year survival rate of patients with high expression of Vimentin protein was 67.44%, while that of patients with low expression of E-cadherin protein was 68.89%. Conclusion: The high expression of Vimentin and low expression of E-cadherin in breast cancer tissues may be related to the occurrence, development, invasion and metastasis of breast cancer. It can be used as a reliable indicator of clinical diagnosis and prognosis.

[Genetic diversity of rhizobial bacteria nodulating Tibetia himalaica in eastern Qinghai-Tibet plateau].

OBJECTIVE: Genetic diversity of the rhizobial bacteria nodulating Tibetia himalaica in the eastern part of Qinghai-Tibet Plateau (Ganzi State, Sichuan) were evaluated. METHODS: The pure culture method was used for isolating the rhiobial strains from the nodules. BOXAIR, 16S rDNA sequences, 16S rDNA PCR-RFLP and Principal Component Analysis (PCA) were used to determine the genetic diversity and phylogenetic relationships. The growth in different salt contents, pH and temperatures were tested. RESULTS: In total 22 strains were isolated from 12 samples in 8 counties. The strains tested were classified into 4 clusters in 16S rDNA PCR-RFLP, and 8 clusters were formed in BOXAIR. The 16S rDNA Simpson genetic diversity index was D = 0. 872. The strains tested were closely related to Rhizobium (11/22 strains), Mesorhizobium (4/22 strains) and Rhizobium- Agrobacterium (7/22 strains). All strains could regularly grow in YMA medium amended with 1% NaCl, and 15/22 strains could grow in 4% NaCl. SCAU679, SCAU694 and SCAU706 could adapt to 7% NaCl, while SCAU689 could tolerant 8% NaCl. Among the strains tested, 15 strains could grow in the pH range from 4.0 to 11; 16 isolates grew well from 4 to 45 degrees C, and all of 22 strains could grow at 28 degrees C after treatment at 60 degrees C for 10 min. CONCLUSION: This study revealed the high genetic diversity of the rhizobia isolated from Tibetia himalaica in the eastern part of Qinghai-Tibet Plateau. The strains tested were adapted to high salt, different range of temperatures and pH.

The radiosensitizing effect of Paeonol on lung adenocarcinoma by augmentation of radiation-induced apoptosis and inhibition of the PI3K/Akt pathway.

PURPOSE: To investigate the radiosensitizing effect and mechanism of action by the natural product Paeonol on lung adenocarcinoma both in vitro and in vivo. MATERIALS AND METHODS: Two lung adenocarcinoma cell lines (human lung adenocarcinoma cell line A549 and mouse Lewis lung carcinoma (LLC) cell line) were chosen for this research. In order to select the experimental concentrations of Paeonol, cytotoxicity was determined using a MTT (3-[4,5-dimethylthiazol-2-yl]-2,5- diphenyltetrazolium bromide) assay. A clonogenic assay was performed to measure the radiosensitizing effects. Apoptosis was determined by the Tunel (terminal deoxynucleotidyl transferase-mediated dUTP nick and labeling) assay and flow cytometry. Protein expression was analyzed by Western blotting. To test the radiosensitizing effect in vivo, a transplanted tumor model was established. RESULTS: The MTT assay showed that Paeonol inhibited proliferation of cells. Paeonol concentration ranged from an IC5 (5% inhibiting concentration) to an IC20 and was used at non-toxic concentrations for subsequent experiments. The clonogenic assay showed that Paeonol enhanced the radiosensitivity of cells. Data from the Tunel assay and flow cytometry verified that Paeonol enhanced radiation-induced apoptosis. Paeonol inhibited the activation of the PI3K/AKT (Phosphatidylinositol 3-kinase/ Protein Kinase B) pathway and down-regulated the expression of COX-2 (Cyclooxygenase-2) and Survivin. Paeonol (1718 mg/kg) combined with 10 Gy irradiation inhibited the growth of a transplanted tumor model in vivo, resulting in the longest tumor growth time, tumor growth delay and the highest inhibition ratio when compared with the radiotherapy alone group. CONCLUSIONS: It is reported for the first time that Paeonol has a radiosensitizing effect on lung adenocarcinoma both in vitro and in vivo. This effect could be related to the augmentation of radiation-induced apoptosis and the inhibition of the PI3K/Akt signalling pathway and its downstream proteins: COX-2 and Survivin.

Synergistic effect of combining paeonol and cisplatin on apoptotic induction of human hepatoma cell lines.

AIM: To investigate whether paeonol (Pae) has synergistic effects with cisplatin (CDDP) on the growth-inhibition and apoptosis-induction of human hepatoma cell lines HepG2 and SMMC-7721. METHODS: The cytotoxic effect of drugs was measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide assay. The coefficient of drug interaction was used to analyze the nature of drug interactions. Morphological changes were observed by acridine orange fluorescence staining. Cell cycle and the apoptosis rate were detected by flow cytometry. Bcl-2 and Bax expression were assayed by immunohistochemical staining. RESULTS: Pae or CDDP had antiproliferative effect on the 2 cell lines in a dose-dependent manner, with different sensitivities to drugs. More interestingly, a synergistic inhibitory effect on the viability of the 2 cell lines was observed after treatment with a combination of Pae (15.63, 31.25, and 62.5 mg/L) with various concentrations of CDDP. Further study showed typical morphological changes of apoptosis if the cells were exposed to the two agents for 24 h. The apoptotic rate of the cells with combination treatment was significantly higher than that of cells treated with Pae or CDDP alone. The expression of Bcl-2 decreased and that of Bax increased in the treated groups, especially in the combination group, with the ratio of Bcl-2/Bax decreasing correspondingly. Additionally, a combination of Pae with CDDP resulted in a stronger S phase arrest, compared with Pae or CDDP alone. CONCLUSION: Pae, in combination with CDDP, had a significantly synergistic growth-inhibitory and apoptosis-inducing effect on the 2 human hepatoma cell lines, which may be useful in hepatoma treatment.

Antiproliferation and apoptosis induction of paeonol in HepG2 cells.

AIM: To investigate the antiproliferative effect of paeonol (Pae) used alone or in combination with chemotherapeutic agents [cisplatin (CDDP), doxorubicin (DOX) and 5-fluorouracil (5-FU)] on human hepatoma cell line HepG2 and the possible mechanisms. METHODS: The cytotoxic effect of drugs on HepG2 cells was measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Morphologic changes were observed by acridine orange (AO) fluorescence staining. Cell cycle and apoptosis rate were detected by flow cytometry (FCM). Drug-drug interactions were analyzed by the coefficient of drug interaction (CDI). RESULTS: Pae (7.81-250 mg/L) had an inhibitory effect on the proliferation of HepG2 cells in a dose-dependent manner, with the IC50 value of (104.77 +/- 7.28) mg/L. AO fluorescence staining and FCM assays showed that Pae induced apoptosis and arrested cell cycle at S phase in HepG2 cells. Further, different extent synergisms were observed when Pae (15.63, 31.25, 62.5 mg/L) was combined with CDDP (0.31-2.5 mg/L), DOX (0.16-1.25 mg/L), or 5-FU (12.5-100 mg/L) at appropriate concentrations. The IC50 value of the three drugs decreased dramatically when combined with Pae (P < 0.01). Of the three different combinations, the sensitivity of cells to drugs was considerably different. CONCLUSION: Pae had a significant growth-inhibitory effect on the human hepatoma cell line HepG2, which may be related to apoptosis induction and cell cycle arrest. It also can enhance the cytotoxicity of chemotherapeutic agents on HepG2 cells, and the S phase arrest induced by Pae may be one of the mechanisms of these interactions.