Efficacy and safety of KL-A167 in previously treated recurrent or metastatic nasopharyngeal carcinoma: a multicenter, single-arm, phase 2 study.

Background: KL-A167 is a fully humanized monoclonal antibody targeting programmed cell death-ligand 1. This phase 2 study aimed to evaluate the efficacy and safety of KL-A167 in Chinese patients with previously treated recurrent or metastatic (R/M) nasopharyngeal carcinoma (NPC). Methods: This was a multicentre, single-arm, phase 2 study of KL-A167 in R/M NPC (KL167-2-05-CTP) (NCT03848286), conducted at 42 hospitals across the People's Republic of China. Eligible patients had histologically confirmed non-keratinising R/M NPC, and had failed at least two lines of chemotherapy. Patients received KL-A167 900mg intravenously once every 2 weeks until confirmed disease progression, intolerable toxicity, or withdrawal of informed consent. The primary endpoint was objective response rate (ORR) assessed by the independent review committee (IRC) according to RECIST v1.1. Findings: Between Feb 26th, 2019 and Jan 13th, 2021, 153 patients were treated. Totally, 132 patients entered full analysis set (FAS) and were evaluated for the efficacy. As of data cutoff date on Jul 13th, 2021, the median follow-up time was 21.7 months (95%CI 19.8-22.5). For FAS population, the IRC-assessed ORR was 26.5% (95%CI 19.2-34.9%), and disease control rate (DCR) was 56.8% (95%CI 47.9-65.4%). Median progression-free survival (PFS) was 2.8 months (95%CI 1.5-4.1) . Median duration of response was 12.4 months (95%CI 6.8-16.5), and median overall survival (OS) was 16.2 months (95%CI 13.4-21.3). When using the cutoff of 1000 copies/ml, 5000 copies/ml and 10,000 copies/ml for plasma EBV DNA titer, baseline low plasma EBV DNA was consistently related with better DCR, PFS and OS. Dynamic change of plasma EBV DNA was significantly associated with ORR and PFS. Among 153 patients, treatment related-adverse events (TRAEs) occurred in 73.2% of patients, and grade ≥3 TRAEs were in 15.0% of patients. No TRAE leading to death was reported. Conclusion: In this study, KL-A167 showed promising efficacy and an acceptable safety profile in patients with previously treated R/M NPC. Baseline plasma EBV DNA copy number might be a potentially useful prognostic biomarker for KL-A167 treatment, and post-treatment EBV DNA decrease might be correlated with better response to KL-A167. Funding: Sichuan Kelun-Biotech Biopharmaceutical Co., Ltd., China National Major Project for New Drug Innovation (2017ZX09304015).

A Simple Technique for Direct Immobilization of Target Enzymes from Cell Lysates Based on the SpyTag/SpyCatcher Spontaneous Reaction.

Many of the current methods for enzyme purification and immobilization suffer from several drawbacks, such as requiring tedious multistep procedures or long preparation, and being environmentally unfriendly, due to the chemicals and conditions involved. Thus, a simple technique for direct purification and immobilization of target enzymes from cell lysates was proposed. The elastin-like polypeptides (ELPs)-SpyCatcher chimera could mediate the formation of silica carriers within seconds and the target enzymes were then covalently immobilized on silica carriers via SpyCatcher/SpyTag spontaneous reaction. These tailor-made carriers were easily prepared, with precisely controlled morphology and size, as well as none-consuming surface modification needed, which could specifically immobilize the SpyTag-fused target enzymes from the cell lysate without pre-purification.

Construction and validation of a prognostic model for stage IIIC endometrial cancer patients after surgery.

BACKGROUND: To explore the most predictive lymph node (LN) scheme for stage IIIC endometrial cancer (EC) patients after hysterectomy and develop a scheme-based nomogram. METHODS: Data from 2626 stage IIIC EC patients, diagnosed between 2010 and 2014, were extracted from the Surveillance, Epidemiology, and End Results (SEER) registry. The predictive ability of four LN schemes was assessed using C-index and Akaike information criterion (AIC). A nomogram based on the most predictive LN scheme was constructed and validated. The comparison of the predictive ability between nomogram and FIGO stage was conducted using the area under the receiver operating characteristic curve (AUC) and decision curve analysis (DCA). RESULTS: FIGO stage (stage IIIC1/stage IIIC2) was not an independent risk factor for OS in stage IIIC EC patients (P = 0.672) and log odds of positive lymph nodes (LODDS) had the best predictive ability (C-index: 0.742; AIC: 8228.95). A nomogram based on LODDS was constructed and validated, which had a decent C-index of 0.742 (0.723-0.762). The nomogram showed a better predictive ability than that of the FIGO staging system. CONCLUSION: FIGO IIIC1/FIGO IIIC2 could not differentiate the prognosis for stage IIIC EC patients. We developed and validated a nomogram based on LODDS to predict OS for post-operative patients with stage IIIC EC.

CHFR­mediated epithelial­to­mesenchymal transition promotes metastasis in human breast cancer cells.

Checkpoint with FHA and RING finger domains (CHFR) is a G2 phase/mitosis checkpoint. Several studies have reported that CHFR is downregulated in multiple cancer types and serves a tumor suppressor role. However, the biological function of CHFR in breast cancer (BRCA), particularly regarding metastasis, are yet to be elucidated. In the present study, it was revealed that CHFR is upregulated in BRCA compared with normal tissues, according to The Cancer Genome Atlas database. In addition, subgroup analysis of BRCA revealed that CHFR was upregulated in both human epidermal growth factor receptor 2­positive and triple­negative BRCA. Meanwhile, patients with high expression levels of CHFR exhibited poorer overall survival rates. Furthermore, the present data revealed that the overexpression of CHFR in SKBR3 cells resulted in enhanced cell migration and invasiveness, and also significantly upregulated mesenchymal markers, such as N­cadherin, vimentin, transcription factor Slug and tight junction protein claudin­1. Furthermore, knockdown of CHFR in MDA­MB­231 cells significantly inhibited cell migration and invasiveness, and also downregulated mesenchymal markers, such as N­cadherin, vimentin and tight junction protein claudin­1. In conclusion, the current results indicated that CHFR expression was associated with cell metastasis in BRCA by mediating epithelial­to­mesenchymal transition.

The efficacy of gefitinib supplementation for breast cancer: A meta-analysis of randomized controlled studies.

INTRODUCTION: The efficacy of gefitinib supplementation for breast cancer remains controversial. We conduct a systematic review and meta-analysis to explore the influence of gefitinib supplementation vs placebo on the efficacy of breast cancer. METHODS: We have searched PubMed, EMbase, Web of science, EBSCO, and Cochrane library databases through February 2019 and included randomized controlled trials assessing the effect of gefitinib supplementation vs placebo on overall response for breast cancer patients. This meta-analysis was performed using the random-effect model. RESULTS: Seven randomized controlled trials involving 927 patients were included in the meta-analysis. Overall, compared with control group for breast cancer, gefitinib supplementation revealed no obvious impact on complete response (risk ration [RR] = 1.19; 95% confidence interval [CI] = 0.58 to 2.44; P = .63), progressive disease (RR = 0.81; 95% CI = 0.59-1.11; P = .18), partial response (RR = 0.67; 95% CI = 0.36-1.25; P = .21), stable disease (RR = 1.02; 95% CI = 0.65-1.60; P = .92), nausea or vomiting (RR = 0.99; 95% CI = 0.73-1.33; P = .93), but was associated with increased incidence of diarrhea (RR = 2.80; 95% CI = 2.23-3.52; P < .00001), decreased incidence of hot flash (RR = 0.53; 95% CI = 0.37-0.78; P = .001), and improved incidence of adverse events (RR = 1.12; 95% CI = 1.05-1.19; P = .0006). CONCLUSIONS: Gefitinib supplementation may provide no positive effect on complete response, progressive disease, partial response or stable disease for breast cancer patients, but with the increase in adverse events.

Fosaprepitant versus aprepitant in the prevention of chemotherapy-induced nausea and vomiting in patients receiving cisplatin-based chemotherapy: a multicenter, randomized, double-blind, double-simulated, positive-controlled phase III trial.

BACKGROUND: To establish the role of antiemetic therapy with neurokinin-1 (NK-1) receptor antagonists (RAs) in Chinese patients associated with cisplatin-base chemotherapy regimens, this study evaluated the efficacy and safety of single-dose intravenous fosaprepitant-based triple antiemetic regimen to a 3-day orally aprepitant-based antiemetic triplet schedule for the prevention of chemotherapy-induced nausea and vomiting (CINV). METHODS: A randomized, double-blind, positive-control design was used to test the noninferiority of fosaprepitant towards aprepitant. Patients receiving cisplatin-base (≥50 mg/m2) chemotherapy were administrated palonosetron and dexamethasone with a single-dose fosaprepitant (150 mg on day 1) or a standard aprepitant regimen (125 mg on day 1, 80 mg on day 2 and day 3). The primary endpoint was complete response (CR) during overall phase (OP). Secondary endpoints include CR during acute phase (AP) and delayed phase (DP), no vomiting and no significant nausea during OP, AP and DP. Accrual of 324 patients per treatment arm was planned to confirm noninferiority with expected CR of 75% and noninferiority margin of minus 10 percentage points. RESULTS: A total of 648 patients were randomly assigned, and 644 were evaluable for efficacy and safety. Antiemetic efficacy of CR during the OP with fosaprepitant and aprepitant was equivalent (71.96% versus 69.35%, P=0.4894). And a between-group difference of 2.61 percentage points was finally achieved (95% CI, -4.42 to 9.64) within predefined bounds for noninferiority (primary end point achieved). Both regimens were well tolerated and commonly reported adverse events (≥1%) were similar between these two group. CONCLUSIONS: Single-dose intravenous fosaprepitant (150 mg) combined with palonosetron and dexamethasone was well tolerated and demonstrated noninferior control of CINV to aprepitant-based triple regimen in Chinese patients treating with cisplatin-base chemotherapy.

The efficacy and safety of panitumumab supplementation for colorectal cancer: A meta-analysis of randomized controlled studies.

BACKGROUND: The efficacy of panitumumab supplementation for colorectal cancer remains controversial. We conduct a systematic review and meta-analysis to explore the influence of panitumumab supplementation on treatment efficacy of colorectal cancer. METHODS: We search PubMed, EMbase, Web of science, EBSCO, and Cochrane library databases through June 2019 for randomized controlled trials (RCTs) assessing the efficacy of panitumumab supplementation for colorectal cancer. This meta-analysis is performed using the random-effect model. RESULTS: Five RCTs are included in the meta-analysis. Overall, compared with control group for colorectal cancer, panitumumab supplementation is associated with the increase in objective response for wild-type (WT) KRAS (RR = 1.70; 95% CI = 1.07-2.69; P = .03), but has no remarkable influence on objective response for mutant KRAS (RR = 0.92; 95% CI = 0.79-1.08; P = .32), objective response (RR = 1.35; 95% CI = 1.00-1.83; P = 0.05), progressive disease for WT KRAS (RR = 0.94; 95% CI = 0.85-1.02; P = .15), mortality (RR = 0.86; 95% CI = 0.69-1.08; P = .20), or mortality for WT KRAS (RR = 0.94; 95% CI = 0.84-1.05; P = .28). In addition, grade 3 and 4 adverse events are found to be higher in panitumumab group than those in control group (RR = 1.17; 95% CI = 1.08-1.27; P = .0001; ). CONCLUSIONS: Panitumumab supplementation can provide some improvement in objective response for colorectal cancer patients with WT KRAS, but results in the increase in grade 3 and 4 adverse events.

Fully Automatic Sliding Motion Compensated and Simultaneous 4D-CBCT via Bilateral Filtering.

PURPOSE: To incorporate the bilateral filtering into the Deformable Vector Field (DVF) based 4D-CBCT reconstruction for realizing a fully automatic sliding motion compensated 4D-CBCT. MATERIALS AND METHODS: Initially, a motion compensated simultaneous algebraic reconstruction technique (mSART) is used to generate a high quality reference phase (e.g. 0% phase) by using all phase projections together with the initial 4D-DVFs. The initial 4D-DVF were generated via Demons registration between 0% phase and each other phase image. The 4D-DVF will then kept updating by matching the forward projection of the deformed high quality 0% phase with the measured projection of the target phase. The loss function during this optimization contains an projection intensity difference matching criterion plus a DVF smoothing constrain term. We introduce a bilateral filtering kernel into the DVF constrain term to estimate the sliding motion automatically. The bilateral filtering kernel contains three sub-kernels: 1) an spatial domain Guassian kernel; 2) an image intensity domain Guassian kernel; and 3) a DVF domain Guassian kernel. By choosing suitable kernel variances, the sliding motion can be extracted. A non-linear conjugate gradient optimizer was used. We validated the algorithm on a non-uniform rotational B-spline based cardiac-torso (NCAT) phantom and four anonymous patient data. For quantification, we used: 1) the Root-Mean-Square-Error (RMSE) together with the Maximum-Error (MaxE); 2) the Dice coefficient of the extracted lung contour from the final reconstructed images and 3) the relative reconstruction error (RE) to evaluate the algorithm's performance. RESULTS: For NCAT phantom, the motion trajectory's RMSE/MaxE are 0.796/1.02 mm for bilateral filtering reconstruction; and 2.704/4.08 mm for original reconstruction. For patient pilot study, the 4D-Dice coefficient obtained with bilateral filtering are consistently higher than that without bilateral filtering. Meantime several image content such as the rib position, the heart edge definition, the fibrous structures all has been better corrected with bilateral filtering. CONCLUSION: We developed a bilateral filtering based fully automatic sliding motion compensated 4D-CBCT scheme. Both digital phantom and initial patient pilot studies confirmed the improved motion estimation and image reconstruction ability. It can be used as a 4D-CBCT image guidance tool for lung SBRT treatment.

Transradial vs transfemoral access in patients with hepatic malignancy and undergoing hepatic interventions: A systematic review and meta-analysis.

BACKGROUND: The aim of this study was to compare transradial access (TRA) approach with transfemoral access (TFA) approach in patients undergoing hepatic interventions. METHODS: We conducted a comprehensive search of the PubMed, Embase, and the Cochrane Library database to identify relevant available articles. Patients' preference, success rate, intra- and postoperative outcomes were analyzed. The risk difference (RD), relative risk (RR), and weighted mean difference (WMD) values were reported with 95% confidence intervals (CIs). We used RevMan 5.3 to perform the pooled analyses. RESULTS: Nine cohort studies were included. A total of 1096 procedures were performed in 877 patients. Of those, 545 procedures (49.7%) were performed by TRA, and 551 procedures (50.3%) were performed by TFA. Patients were significantly prefer the TRA (86.5%) to the TFA (13.5%) (RD = 0.88, P < .00001). The procedure time in TRA groups was longer (WMD = 3.36, 95% CI 1.24-5.47, P = .002). But there were no significant difference in terms of success rate, fluoroscopy time, radiation dosage, contrast volume, and postoperative vascular complications. CONCLUSION: For patients suffer from primary or secondary hepatic malignancy and undergoing hepatic interventions, the present meta-analysis demonstrated that patients prefer the TRA approach to the TFA approach. But the procedure time is longer in TRA group.

Krüppel-like factor 8 regulates VEGFA expression and angiogenesis in hepatocellular carcinoma.

Tumor angiogenesis plays a critical role in hepatocellular carcinoma (HCC) development and progression, but its mechanism is unclear. Krüppel-like factor 8 (KLF8) is a transcription factor that plays an important role in HCC progression. Here, we investigated the role of KLF8 in angiogenesis in HCC and its possible mechanism. Immunohistochemistry, quantitative RT-PCR, western blotting, promoter reporter assays, chromatin immunoprecipitation (ChIP), and chicken chorioallantoic membrane (CAM) and nude mouse tumor models were used to show that the mRNA and protein expression levels of KLF8 and VEGFA are highly correlated in HCC tissue samples. The up-regulation of KLF8 increased VEGFA protein levels and induced VEGFA promoter activity by binding to the CACCC region of the VEGFA promoter. In addition, KLF8 regulated HIF-1α and Focal adhesion kinase (FAK) expression. The PI3K/AKT inhibitor LY294002 inhibited KLF8-induced VEGFA expression, whereas PI3K/AKT signaling pathway proteins, such as P-PDK1(Ser241) and P-AKT(Thr308), were decreased significantly. KLF8-overexpressing HCC cells had a higher potential for inducing angiogenesis. Thus, our results indicate that KLF8 may induce angiogenesis in HCC by binding to the CACCC region of the VEGFA promoter to induce VEGFA promoter activity and through FAK to activate PI3K/AKT signaling to regulate HIF-1α expression levels.

Triplet Chemotherapy (FOLFOXIRI) Plus Bevacizumab Versus Doublet Chemotherapy (FOLFOX/FOLFIRI) Plus Bevacizumab in Conversion Therapy for Metastatic Colorectal Cancer: a Meta-Analysis.

BACKGROUND/AIMS: Conversion therapy can convert unresectable metastatic colorectal cancer (mCRC) into resectable. However, the optimal conversion regimen was not yet defined. This meta-analysis aimed to compare the efficacy and safety of the triplet chemotherapy (FOLFOXIRI) plus bevacizumab (Bev) with doublet chemotherapy (FOLFOX/FOLFIRI) plus Bev in conversion therapy. METHODS: Randomized controlled trials (RCTs) from databases, including Pubmed, EMBASE, Cochrane clinical trials, clinicaltrial.gov and some conferences, were searched from the inception to November 2017. The R0 resection, objective response rate (ORR), progression-free survival (PFS), overall survival (OS) and the incidence of adverse events were pooled with the use of hazard ratio (HR) or risk ratio (RR). RESULTS: Four RCTs with 1013 patients were included. FOLFOXIRI plus Bev regimen significantly improved the overall R0 resection rate (RR 1.41, 95% confidence interval (CI) 1.07-1.85, I2=37%), liver R0 resection rate (RR 2.28, 95% CI 1.34-3.89, I2=0%), ORR (RR 1.20, 95% CI 1.09-1.32, I2=0%), PFS (HR 0.72, 95% CI 0.62-0.84, I2=36%) and OS (HR 0.80, 95% CI 0.66-0.97, I2=0%). There was no significant difference in any Grade≥3 adverse event (RR 1.08, 95% CI 0.99-1.17, I2=0%) between two regimens. FOLFOXIRI-Bev was associated with a higher risk of neutropenia (RR 1.77, 95% CI 1.13-2.79, I2=68%) and diarrhea (RR 1.65, 95% CI 1.17-2.32, I2=0%). CONCLUSIONS: Triplet chemotherapy plus Bev significantly improved the R0 resection rates, ORR, PFS and OS in comparison with doublet chemotherapy plus Bev in conversion therapy for mCRC patients, with a higher risk of neutropenia and diarrhea.

Bevacizumab combined with chemotherapy for ovarian cancer: an updated systematic review and meta-analysis of randomized controlled trials.

BACKGROUND: This meta-analysis was updated with results from a new trial and final data to reassess the efficacy and safety of bevacizumab combined with chemotherapy in ovarian cancer (OC). METHODS: Randomized controlled trials (RCTs) were searched in PubMed, EMBASE, Cochrane clinical trials, Web of Science and clinicaltrial.gov databases. Outcomes included the progression-free survival (PFS), overall survival (OS), objective response rate (ORR) and common adverse events. The hazard ratio (HR), risk ratio (RR) and odds ratio (OR) were pooled when the meta-analysis was performed. RESULTS: Five RCTs with 4994 patients were included. In overall newly diagnosed OC, bevacizumab combined with chemotherapy did not significantly improve PFS (HR 0.85, 95%CI 0.70-1.02) or OS (HR 0.94, 95%CI 0.84-1.05). In the high-risk progression subgroup, the addition of bevacizumab significantly improved PFS (HR 0.76, 95%CI 0.68-0.84) and OS (HR 0.85, 95%CI 0.74-0.96). In recurrent OC, the addition of bevacizumab to chemotherapy significantly extended PFS (HR 0.53, 95%CI 0.45-0.63) and OS (HR 0.87, 95%CI 0.77-0.99). The ORR was improved (OR 2.37, 95%CI 1.99-2.82) in the overall population. Bevacizumab increased the incidence of hypertension (RR 21.27, 95%CI 9.42-48.02), proteinuria (RR 4.77, 95%CI 2.15-10.61), bleeding (RR 3.16, 95%CI 1.59-6.30), GI perforations (RR 2.76, 95%CI 1.51-5.03), arterial thrombosis events (RR 2.39, 95%CI 1.39-4.10) and venous thrombosis events (RR 1.43, 95%CI 1.04-1.96). CONCLUSIONS: Bevacizumab combined with chemotherapy significantly improved PFS and OS in both patients with high-risk of progression and patients with recurrent OC, with an increased incidence of common adverse events. However, no statistically significant survival benefit was identified in the front-line settings.

Predictive parameters for selection of electronic tissue compensation radiotherapy in early-stage breast cancer patients after breast-conserving surgery.

Electronic tissue compensation (eComp) is an external beam planning technique allowing user to manually generate dynamic beam fluence to produce more uniform or modulated dose distribution. In this study, we compared the effectiveness between conventional three-dimensional conformal radiotherapy (3DCRT) and eComp for whole breast irradiation. 3DCRT and eComp planning techniques were used to generate treatment plans for 60 whole breast patients, respectively. The planning goal was to cover 95% of the planning target volume (PTV) with 95% of the prescription dose while minimizing doses to lung, heart, and skin. Comparing to 3DCRT plans, on the average, eComp treatment planning process was about 7 minutes longer, but resulted in lower lung V20Gy, lower mean skin dose, with similar heart dose. The benefits were more pronounced for larger breast patients. Statistical analyses were performed between critical organ doses and patient anatomic features, i.e., central lung distance (CLD), maximal heart distance (MHD), maximal heart length (MHL) and breast separation (BS) to explore any correlations and planning method selection. It was found that to keep the lung V20Gy lower than 20% and mean skin dose lower than 85% of the prescription dose, eComp was the preferred method for patients with more than 2.3 cm CLD or larger than 22.5 cm BS. The study results may be useful in providing a handy criterion in clinical practice allowing us to easily choose between different planning techniques to satisfy the planning goal with minimal increase in complexity and cost.

Association between vitamin D receptor gene polymorphisms and breast cancer in a Chinese population.

OBJECTIVE: The present study aimed to explore the association between vitamin D receptor (VDR) genetic polymorphisms and breast cancer risk. METHODS: A total of 219 patients with breast cancer and 321 cases of females without breast cancer were enrolled for the present study. PCR-RFLP method was used to genotype 3 SNPs of the VDR gene (rs1544410, rs7975232 and rs731236). ELISA method was used to detect the amount of 1,25(OH)2D3 in the plasma. The results were analyzed using SPSS 17.0 software. RESULTS: We found rs7975232 was associated with breast cancer risk. Compared with GG genotype carriers, TT subjects had a lower risk of breast cancer (P = 0. 004, OR = 0.774, 95% CI: 0.212~0.955). However, there was no difference in 1,25(OH)2D3 levels among the different genotypes. In addition, the distribution frequency of the haplotype A-G-T in the patients group was 4.4%, significantly higher than the control group (0.7%, P = 0.003; OR=2.643, 95% CI: 1.631~7.012), while the distribution of haplotype G-T-T in the patient group was significantly lower than in the control group (17.3% vs. 28.4%, P = 0.011, OR = 0.543, 95% CI: 0.325~0.854). CONCLUSIONS: The VDR gene was associated with breast cancer pathogenesis; females carrying the haplotype G-T-T had a lower breast cancer risk, while the haplotype A-G-T conferred a higher risk.

[Combined inhibition of PI3K and MEK has synergistic inhibitory effect on the proliferation of cisplatin-resistant ovarian cancer cells].

OBJECTIVE: To investigate the effect of phosphatidylinositol 3 kinase (PI3K) inhibitor LY294002 combined with mitogen activated protein kinase kinase (MEK) inhibitor AZD6244 on the proliferation of cisplatin-resistant SKOV3/DDP ovarian cancer cell line. METHODS: The alteration in the cell proliferation of SKOV3/DDP cells treated with 5, 10, 20, 40, 80 µmol/L LY294002 and 1, 2, 4, 8, 16 µmol/L AZD6244 alone or together, was detected by MTT assay. The 50% inhibitory concentration (IC50) and combined index (CI) were calculated. The concentration of the combination was obtained based on the MTT assay, and the cells were divided into four groups: the control group, LY294002 group (5 µmol/L), AZD6244 group (7 µmol/L) and combination group (LY294002 5 µmol/L and AZD6244 7 µmol/L); Forty-eight hours later, MTT assay was used to detect the cell proliferation and calculate cell doubling time; colony formation assay was performed to detect colony formation efficiency; Annexin V-PE/7-ADD staining combined with flow cytometry (FCM) was used to detect the apoptotic rates and cell cycle; Western blotting was employed to detect the levels of AKT, phosphorylated AKT (p-AKT), extracellular signal-regulated kinase 1/2 (ERK1/2), phosphorylated ERK1/2 (p-ERK1/2), cyclin D1 and cleaved caspase-3 protein. RESULTS: Cell growth was inhibited by LY294002 or AZD6244 alone, and the effect was strengthened when LY294002 was combined with AZD6244 (CI<1), indicating that the two inhibitors showed synergistic effect. The proliferation of cells was significantly slower, the cell doubling time was significantly prolonged, and colony number was reduced in the combined treatment group (P<0.01) compared with the single inhibitor treatment group and the control group. Meanwhile, FCM demonstrated that the apoptotic rate of the combination group was significantly higher than that of the other groups, and the cells significantly increased in G1 phase and decreased in S phase (P<0.05). As Western blotting showed, there were no differences in the expressions of AKT and ERK1/2 protein among the four groups (P>0.05); in the LY294002 treatment group, the level of p-AKT protein decreased and p-ERK increased; in the AZD6244 group, the level of p-ERK1/2 protein decreased and p-AKT increased; in the combination group, the levels of p-AKT, p-ERK1/2 and cyclin D1 protein were significantly inhibited, while cleaved caspase-3 protein was up-regulated. CONCLUSION: Combined PI3K inhibitor LY294002 and MEK inhibitor AZD6244 has synergistic effect on inhibition of SKOV3/DDP cell growth by inducing apoptosis and blocking cell cycle.

[Inhibitive effect of celecoxib combined with tegafur gimeracil oteracil potassium on the growth of xenograft tumor of gastric cancer in nude mice].

OBJECTIVE: To investigate the anti-tumor effect of celecoxib combined with tegafur gimeracil oteracil potassium on subcutaneous xenograft tumor of gastric cancer in nude mice and analyze the possible mechanism. METHODS: A xenograft tumor model of gastric cancer was established subcutaneously in nude mice. After the largest diameter of tumor reached about 5 mm, the nude mice were randomly divided into 4 groups: the control group, the celecoxib group, the tegafur gimeracil oteracil potassium group, and the combination group; the drug was administered respectively for 21 days. Thereafter, tumor tissues were collected, tumor volume was measured, and tumor inhibition rate was calculated. Apoptosis was determined by TUNEL assay and the expression levels of PCNA, Bcl-2 and caspase-3 by immunohistochemistry. RESULTS: The tumor inhibition rates of the celecoxib group, the tegafur gimeracil oteracil potassium group, the combination group were 30.8%, 50.1%, 78.8%, respectively. The apoptosis index in treatment groups was higher than that in the control group (P<0.01), and the combination group was higher than single drug group (P<0.01). The expressions of PCNA, Bcl-2 in treatment groups were lower than those in the control group (P<0.01), and the combination group was lower than single drug group (P<0.05). The expression of caspase-3 in treatment groups was higher than that in the control group (P<0.05), and the combination group was higher than single drug group (P<0.01). CONCLUSION: Both celecoxib and tegafur gimeracil oteracil potassium showed obvious anti-tumor effect, and the combination of the two acted synergistically. The possible mechanism was that they inhibited tumor growth through inhibiting proliferation and promoting apoptosis of the tumor cells.

Suramin inhibits the growth of nasopharyngeal carcinoma cells via the downregulation of osteopontin.

Radiotherapy is the principal therapy for nasopharyngeal carcinoma (NPC) at early stages. A number of chemotherapeutic methods have been used to inhibit the progression of NPC at elevated stages. Suramin has been reported to inhibit the growth of certain tumor cells via various pathways. In the present study, we aimed to analyze the effects of suramin on the proliferation of NPC cells (CNE-2). Suramin was proved to demonstrate NPC cell growth-inhibiting effects both in a dose- and time-dependent manner. To determine the potential mechanisms of these effects, western blotting and flow cytometric analysis were performed. Suramin was found to have the potential to induce cell cycle arrest in S-phase CNE-2 cells. Additionally, we found that the OPN level may decrease in suramin-treated CNE-2 cells. The changes of certain apoptosis- and p-AKT-associated proteins possibly regulating the OPN expression were measured by western blotting. In conclusion, suramin may function as a potential agent for the adjunctive therapy of NPC.

[Relationship between the radiosensitivity of hepatic carcinoma cells and their survivin expression levels].

OBJECTIVE: To investigate the relationship between the radiosensitivity of hepatic carcinoma cells and their survivin expression levels. METHODS: Hepatic carcinoma cell lines HepG2 and SMMC-7721 were irradiated with various doses of 60Co gamma-rays. The cell survival rate, expression of survivin, cell cycle profile and activity of caspase-3 were respectively detected by clonogenic assay, immunocytochemistry, flow cytometry and chromatometry. RESULTS: The surviving fraction at 2Gy (SF2) of HepG2 and SMMC-7721 were 0.43+/-0.01 vs 0.70+/-0.02, and HepG2 had higher radiosensitivity than SMMC-7721. gamma-rays radiation up-regulated the expression of survivin. SMMC-7721 had a significantly higher expression of survivin than HepG2 (t = 2.81-5.20, P < 0.05). The activity of caspase-3 was more powerful in HepG2 than in SMMC-7721 (t = 6.05-6.72, P < 0.01). CONCLUSION: Survivin may play a critical role in mediating radiation resistance in SMMC-7721 through its up-regulation and caspase-3 dependent manner.

Detecting normal cell radiosensitivity via assay of DNA damage in lymphocytes for individualizing radiotherapy in head and neck cancer patients.

PURPOSE: The purpose of this study was to determine whether the distribution of radiosensitivities in normal tissues of head and neck cancer patients, measured using a DNA damage assay on lymphocytes, is likely to provide sufficient discrimination to enable reliable identification of patients with abnormal sensitivities. MATERIAL AND METHODS: Radiosensitivity was assessed in 307 lymphocyte samples from unselected head and neck cancer patients and was quantified as the initial number of DNA double-strand breaks (dsb) induced per Gray and per DNA unit (200 Mbp). RESULTS: The existence of an inter-individual variation in the radiosensitivity parameter is described by the range (0.41--9.38 dsb/Gy/DNA unit) of the values found. We detected 37 patients who developed severe skin reactions during radiotherapy treatment and we compared their radiosensitivity values with the remaining patients treated. Radiosensitivity values of >7.20 dsb/Gy/DNA unit should theoretically correspond to highly radiosensitive patients. CONCLUSIONS: Our results suggest that initial DNA damage measured on lymphocytes offers an approach to predict the acute response of human normal tissues prior to radiotherapy. .