Effect of Raltitrexed on ECA109 Cellular Radiosensitivity and Its Mechanism in Esophageal Cancer.

BACKGROUND: To investigate the effect of raltitrexed + X-ray irradiation on esophageal cancer ECA109 cells and analyze the potential action mechanism. METHODS: The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was used to analyze the inhibitory effect of raltitrexed on cell proliferation. The effect of raltitrexed on radiosensitivity was studied through a clone-forming experiment. The scratch assay and invasion test were performed to understand the cell migration and invasion abilities. The apoptosis rate change was measured using a flow cytometer, and Western Blotting was used to determine the expression of B cell lymphoma-2 (Bcl-2) and Bcl2-associated X protein (Bax) in each group. RESULTS: Raltitrexed significantly inhibited ECA109 proliferation in a time-dose-dependent manner; there were significant differences among different concentrations and times of action. The results of the clone-forming experiment showed a sensitization enhancement ratio of 1.65, and this demonstrated a radiosensitization effect. After the combination of raltitrexed with X-ray, the cell migration distance was shortened, and the number of cells penetrating the membrane was reduced. CONCLUSION: Raltitrexed can inhibit the growth of esophageal cancer ECA109 cells and has a radiosensitization effect.

Microbiome profiling and Co-metabolism pathway analysis in cervical cancer patients with acute radiation enteritis.

Background: Intestinal bacteria significantly contribute to the metabolism of intestinal epithelial tissues. As the occurrence and development of radiation enteritis (RE) depend on the "co-metabolism" microenvironment formed by the host and intestinal microbiota, which involves complex influencing factors and strong correlations, ordinary techniques struggle to fully explain the underlying mechanisms. However, given that it is based on systems biology, metabolomics analysis is well-suited to address these issues. This study aimed to analyze the metabolomic changes in urine, serum, and fecal samples during volumetric modulated arc therapy (VMAT) for cervical cancer and screen for characteristic metabolites of severe acute radiation enteritis (SARE) and RE. Methods: We enrolled 50 patients who received radiotherapy for cervical cancer. Urine, serum, and fecal samples of patients were collected at one day before radiotherapy and the second week, fourth week, and sixth week after the start of radiotherapy. Control group samples were collected during the baseline period. Differential metabolites were identified by metabolomics analysis; co-metabolic pathways were clarified. We used the mini-SOM library for incorporating characteristic metabolites, and established metabolite classification models for predicting SARE and RE. Results: Urine and serum sample data showed remarkable clustering effect; metabolomics data of the fecal supernatant were evidently disturbed. Patient sample analyses during VMAT revealed the following. Urine samples: Downregulation of the pyrimidine and riboflavin metabolism pathways as well as initial upregulation followed by downregulation of arginine and proline metabolism pathways and the arginine biosynthesis pathway. Fecal samples: Upregulation of linoleic acid and phenylalanine metabolic pathways and initial downregulation followed by upregulation of arachidonic acid (AA) metabolic pathways. Serum samples: Initial upregulation followed by downregulation of the arginine biosynthesis pathway and downregulation of glutathione, AA, and arginine and proline metabolic pathways. Conclusion: Patients with cervical cancer exhibited characteristic metabolic pathways and characteristic metabolites predicting RE and SARE were screened out. An effective RE mini-SOM classification model was successfully established.

Establishment of a prediction model for severe acute radiation enteritis associated with cervical cancer radiotherapy.

BACKGROUND: Cervical cancer is one of the most common gynecological malignant tumors. Radiation enteritis (RE) leads to radiotherapy intolerance or termination of radiotherapy, which negatively impacts the therapeutic effect and seriously affects the quality of life of patients. If the incidence of RE in patients can be predicted in advance, and targeted clinical preventive treatment can be carried out, the side effects of radiotherapy in cervical cancer patients can be significantly reduced. Furthermore, accurate prediction of RE is essential for the selection of individualized radiation dose and the optimization of the radiotherapy plan. AIM: To analyze the relationships between severe acute RE (SARE) of cervical cancer radiotherapy and clinical factors and dose-volume parameters retrospectively. METHODS: We included 50 cervical cancer patients who received volumetric modulated arc therapy (VMAT) from September 2017 to June 2018 in the Department of Radiotherapy at The First Affiliated Hospital Soochow University. Clinical and dose-volume histogram factors of patients were collected. Logistic regression analysis was used to evaluate the predictive value of each factor for SARE. A nomogram to predict SARE was developed (SARE scoring system ≥ 3 points) based on the multiple regression coefficients; validity was verified by an internal verification method. RESULTS: Gastrointestinal and hematological toxicity of cervical cancer VMAT gradually increased with radiotherapy and reached the peak at the end of radiotherapy. The main adverse reactions were diarrhea, abdominal pain, colitis, anal swelling, and blood in the stool. There was no significant difference in the incidence of gastrointestinal toxicity between the radical and postoperative adjuvant radiotherapy groups (P > 0.05). There were significant differences in the small intestine V20, V30, V40, and rectal V40 between adjuvant radiotherapy and radical radiotherapy after surgery (P < 0.05). Univariate and multivariate analyses revealed anal bulge rating (OR: 14.779, 95%CI: 1.281-170.547, P = 0.031) and disease activity index (DAI) score (OR: 53.928, 95%CI: 3.822-760.948, P = 0.003) as independent predictors of SARE. CONCLUSION: Anal bulge rating (> 0.500 grade) and DAI score (> 2.165 points) can predict SARE. The nomogram shows potential value in clinical practice.

Effects and mechanisms of GSG2 in esophageal cancer progression.

BACKGROUND: Esophageal cancer was recognized as one of the malignant tumors with poor prognosis. Germ cell associated 2 (GSG2) has been reported to be of great significance in cell growth and tumor formation. This study aimed to investigate the biological function and molecular mechanism of GSG2 in esophageal cancer. METHODS: First, relationship between GSG2 expression and tumor characteristics in esophageal cancer patients was analyzed through immunohistochemical (IHC) staining. MTT assay, flow cytometry, cloning formation assay, wound-healing assay and Transwell assay were used to determine proliferation, apoptosis and migration of esophageal cancer cell with GSG2 knockdown in vitro. Expression of apoptosis related proteins and downstream pathway proteins after GSG2 knockdown were detected through Human Apoptosis Antibody Array and western blot analysis. The GSG2 knockdown function in vivo was explored through a xenograft tumor model. RESULTS: GSG2 was highly expressed in tumor tissues, which has clinical significance in predicting the malignant degree of patients with esophageal cancer. In addition, GSG2 knockdown significantly inhibited a variety of malignant biological behaviors of esophageal cancer cells, such as inhibiting proliferation, reducing colony formation, promoting apoptosis, hindering migration. The decrease of GSG2 expression in esophageal cancer cells can inhibit the xenograft tumor growth. CONCLUSIONS: In conclusion, GSG2 was involved in esophageal cancer progression and development, which may provide an effective molecular target for the treatment of esophageal cancer in the future.

Analysis of nutritional risk, skeletal muscle depletion, and lipid metabolism phenotype in acute radiation enteritis.

BACKGROUND: Radiation enteritis, which often occurs during radiation-induced acute intestinal symptoms (RIAIS), is the most common and important complication during radiotherapy for cervical cancer. RIAIS caused by abdominal and pelvic radiotherapy will affect nutrient intake, digestion, absorption, and metabolism, leading to malnutrition or poorer nutritional status. In patients with malignant tumors, malnutrition can adversely affect the curative effect and response of radiotherapy by reducing radiosensitivity, affecting the precision of radiotherapy placement and increasing the incidence of radiotherapy-related adverse reactions. AIM: To analyze nutritional risk, skeletal muscle depletion, and lipid metabolism phenotype in acute radiation enteritis. METHODS: Fifty patients with cervical cancer received external beam radiotherapy, and 15 patients received brachytherapy after external beam radiotherapy. Body weight, body composition parameters, nutritional risk screening (NRS) 2002 score, and blood biochemical indices of patients with cervical cancer during periradiation were tested by a one-way repeated measures analysis of variance. Metabolomics analysis was used to identify characteristic lipid metabolism pathways. Clinical factors that affect linoleic acid changes were screened using the generalized evaluation equation. RESULTS: Among the 50 patients, 37 had RIAIS, including 34 patients with grade 1-2 RIAIS and 3 patients with grade 3 RIAIS. The NRS 2002 score of patients who underwent cervical cancer radiotherapy continued to increase during the periradiation period, and 42 patients who underwent cancer radiotherapy had nutritional deficits (NRS 2002 score ≥ 3 points) at the end of radiotherapy. Correlation analyses revealed that body weight and body mass index changes were closely associated with body fat content (R2 = 0.64/0.51). The results of the univariate analysis showed that radiotherapy time, percentage reduction of serum albumin, and percentage reduction of serum prealbumin were the key factors affecting skeletal muscle exhaustion (P < 0.05). Metabolomic analysis of fecal supernatants of cervical cancer patients during the periradiation period revealed the involvement of linoleic acid, cholic acid, arachidonic acid, and N-acetyl-L-benzene alanine in the metabolic pathway of linoleic acid. CONCLUSION: Cervical cancer radiotherapy patients faced nutritional risks, decreased serum albumin synthesis, and increased risk of skeletal muscle exhaustion. Linoleic acid was a biomarker of high nutritional risk.

Clinical evaluation of deep learning-based clinical target volume three-channel auto-segmentation algorithm for adaptive radiotherapy in cervical cancer.

OBJECTIVES: Accurate contouring of the clinical target volume (CTV) is a key element of radiotherapy in cervical cancer. We validated a novel deep learning (DL)-based auto-segmentation algorithm for CTVs in cervical cancer called the three-channel adaptive auto-segmentation network (TCAS). METHODS: A total of 107 cases were collected and contoured by senior radiation oncologists (ROs). Each case consisted of the following: (1) contrast-enhanced CT scan for positioning, (2) the related CTV, (3) multiple plain CT scans during treatment and (4) the related CTV. After registration between (1) and (3) for the same patient, the aligned image and CTV were generated. Method 1 is rigid registration, method 2 is deformable registration, and the aligned CTV is seen as the result. Method 3 is rigid registration and TCAS, method 4 is deformable registration and TCAS, and the result is generated by a DL-based method. RESULTS: From the 107 cases, 15 pairs were selected as the test set. The dice similarity coefficient (DSC) of method 1 was 0.8155 ± 0.0368; the DSC of method 2 was 0.8277 ± 0.0315; the DSCs of method 3 and 4 were 0.8914 ± 0.0294 and 0.8921 ± 0.0231, respectively. The mean surface distance and Hausdorff distance of methods 3 and 4 were markedly better than those of method 1 and 2. CONCLUSIONS: The TCAS achieved comparable accuracy to the manual delineation performed by senior ROs and was significantly better than direct registration.

Individualized Whole Course Nutrition Management for Nasopharyngeal Carcinoma Patients Undergoing Radiotherapy.

BACKGROUND: Radiotherapy-induced oral mucositis (RIOM) is the most common toxicity associated with radiotherapy for nasopharyngeal carcinoma (NPC). Patients with RIOM become malnourished, which can affect the delivery and dose of radiotherapy. The value of personalizing nutrition recommendations for cancer prevention and management is increasingly recognized. To investigate the effect of individualized whole course nutrition management on nutritional status and the incidence and severity of RIOM in NPCs. METHODS: This retrospective study included 77 patients who were provided individualized whole course nutrition management during radiotherapy (RT) and a 1-month follow-up. Seventy-one patients were included in the control group. RESULTS: During radiotherapy, severity of RIOM was significantly lower in the intervention group. There were statistically significant differences in oral mucosa recovery time and nutritional status between the two groups (p < 0.05). CONCLUSIONS: Individualized whole course nutrition management had the potential to maintain nutritional status and decrease the adverse effects of radiotherapy in NPCs.

Deep learning-based auto-segmentation of clinical target volumes for radiotherapy treatment of cervical cancer.

OBJECTIVES: Because radiotherapy is indispensible for treating cervical cancer, it is critical to accurately and efficiently delineate the radiation targets. We evaluated a deep learning (DL)-based auto-segmentation algorithm for automatic contouring of clinical target volumes (CTVs) in cervical cancers. METHODS: Computed tomography (CT) datasets from 535 cervical cancers treated with definitive or postoperative radiotherapy were collected. A DL tool based on VB-Net was developed to delineate CTVs of the pelvic lymph drainage area (dCTV1) and parametrial area (dCTV2) in the definitive radiotherapy group. The training/validation/test number is 157/20/23. CTV of the pelvic lymph drainage area (pCTV1) was delineated in the postoperative radiotherapy group. The training/validation/test number is 272/30/33. Dice similarity coefficient (DSC), mean surface distance (MSD), and Hausdorff distance (HD) were used to evaluate the contouring accuracy. Contouring times were recorded for efficiency comparison. RESULTS: The mean DSC, MSD, and HD values for our DL-based tool were 0.88/1.32 mm/21.60 mm for dCTV1, 0.70/2.42 mm/22.44 mm for dCTV2, and 0.86/1.15 mm/20.78 mm for pCTV1. Only minor modifications were needed for 63.5% of auto-segmentations to meet the clinical requirements. The contouring accuracy of the DL-based tool was comparable to that of senior radiation oncologists and was superior to that of junior/intermediate radiation oncologists. Additionally, DL assistance improved the performance of junior radiation oncologists for dCTV2 and pCTV1 contouring (mean DSC increases: 0.20 for dCTV2, 0.03 for pCTV1; mean contouring time decrease: 9.8 min for dCTV2, 28.9 min for pCTV1). CONCLUSIONS: DL-based auto-segmentation improves CTV contouring accuracy, reduces contouring time, and improves clinical efficiency for treating cervical cancer.

Expression and clinicopathological significance of Foxp3 and VISTA in cervical cancer.

OBJECTIVE: To detect the expression differences of Foxp3 and VISTA in chronic cervical inflammation, cervical intraepithelial neoplasia, and cervical cancer, and to explore the role of Foxp3 and VISTA in the development of cervical cancer and the effect of Foxp3 and VISTA on the prognosis of cervical cancer, to provide a theoretical basis for clinical immunotherapy of cervical cancer. METHODS: We collected 130 paraffin specimens of cervical tissue, which included 70 cases of cervical cancer tissue, 40 cases of cervical intraepithelial neoplasia tissues and 20 cases of chronic cervicitis. The expression of Foxp3 and VISTA in each group was detected, and the study was conducted based on the clinicopathological characteristics of the patients. The patients were followed up and the prognosis was statistically analyzed. RESULT: 1. The expression of Foxp3 and VISTA was statistically different between the cervical cancer group and other groups. 2. Expressions of Foxp3 and VISTA were significantly correlated. 3. In 70 cases of cervical cancer, the expression of Foxp3 and VISTA was related to the clinical stage. 4. The 3-year survival rate of 70 patients with cervical cancer was 72.9%, and there were no factors affecting 3-year OS found. The expression of Foxp3 and VISTA was significantly correlated with the prognosis of cervical cancer. Foxp3 and VISTA double positive expression group had the worst prognosis. CONCLUSION: 1. In cervical cancer, the expression of Foxp3 and VISTA was significantly higher than that of cervical intraepithelial neoplasia and chronic cervicitis, which suggested that they were closely related to the occurrence and growth of cervical cancer. 2. The expression of Foxp3 and VISTA was significantly related. 3. The positive expression of Foxp3 and VISTA could be used as independent prognostic factors for cervical cancer prognosis providing a strong basis for cervical cancer immunotherapy.

lncRNA LINC00460 Functions as a Competing Endogenous RNA and Regulates Expression of BGN by Sponging miR-149-5p in Colorectal Cancer.

BACKGROUND AND AIM: There are an increasing number of studies indicating the important roles served by long non-coding RNAs (lncRNAs) in the development of different types of cancer. LINC00460 is a novel identified lncRNA that was found to be upregulated in colorectal cancer. However, the biological roles of LINC00460 in colorectal cancer have yet to be fully elucidated. This study was aimed to investigate the functions and molecular mechanisms of LINC00460 on colorectal cancer metastasis. METHODS: Expression of LINC00460 and biglycan (BGN) in colorectal cancer tissues and cell lines were quantified by real time PCR or western blotting assay. Cell migration and invasion assays were performed to determine the effect of LINC00460 on tumor metastasis in vitro. The binding interaction between microRNA-149-5p and LINC00460 was revealed by luciferase reporter assay. RESULTS: In the present study, lncRNA LINC00460 was shown to be upregulated in colorectal cancer tissues, and overexpression of LINC00460 significantly promoted metastasis of colorectal cancer in vitro. Furthermore, miR-149-5p interacted with LINC00460, and they negatively regulated expression of each other. Transfection of miR-149-5p mimics partially counteracted the tumor metastasis-promoting effects induced by LINC00460 overexpression. Finally, overexpression of LINC00460 upregulated the expression levels of biglycan, a target gene of miR-149-5p, which has also been identified as an oncogenic driver in colorectal cancer. CONCLUSION: Taken together, the present study demonstrated that LINC00460 promoted metastasis of CRC by sponging miR-149-5p and thereby affecting biglycan expression levels.

The Combination of Preoperative Nutritional Risk Screening-2002 and Neutrophil-to-Lymphocyte Ratio is a Useful Prognostic Marker in Patients with Esophageal Squamous Cell Carcinoma.

The aim of this study was to investigate the prognostic values of a novel evaluated system, named the CONN (combination of Nutritional Risk Screening 2002 [NRS-2002] and neutrophil-to-lymphocyte ratio [NLR]), in patients with esophageal squamous cell carcinoma (ESCC) by curative esophagectomy. A total of 278 patients with ESCC receiving standard curative esophagectomy were retrospectively analyzed. The CONN was calculated by combined NRS-2002 and NLR according to the corresponding cutoff values: patients with both elevated NRS-2002 (≥3.0) and NLR (≥3.0) were allocated a score of 2, and patients showing one or neither were allocated a score of 1 or 0, respectively. In our univariate analysis, the following factors were significantly associated with poor PFS and OS: T stage, N stage, TNM stage, NLR, NRS-2002 and CONN (all P < 0.05). Furthermore, multivariate Cox regression analysis showed that N stage (P = 0.039), NRS-2002 (P = 0.041) and CONN (P = 0.001) were independent prognostic factors for PFS. While T stage (P = 0.017), N stage (P = 0.048), NLR (P = 0.021), NRS-2002 (P = 0.001) and CONN (P = 0.001) were independent prognostic factors for OS. In conclusion, CONN was an independent prognostic marker for survival prediction in patients with ESCC receiving surgery.

Whole-Brain Radiotherapy Can Improve the Survival of Patients with Multiple Brain Metastases from Non-Small Cell Lung Cancer Treated by Epidermal Growth Factor Receptor-Tyrosine Kinase Inhibitors.

OBJECTIVE: To observe whether whole-brain radiotherapy (WBRT) can bring survival benefits to patients with multiple brain metastases (BM) from non-small cell lung cancer (NSCLC) treated by epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor (TKI) and determine the best time for WBRT intervention. METHODS: A retrospective analysis was performed on 148 patients diagnosed with EGFR gene-mutated NSCLC. All patients had multiple BM and received EGFR-TKI targeted therapy, which was performed to observe whether WBRT can bring survival benefits, and whether the choice of WBRT timing affects the survival of patients. RESULTS: Among the 148 patients with NSCLC treated with EGFR-TKI, 76 received WBRT; 72 were without WBRT. WBRT can reduce the intracranial progression rate in the patients (19.7% vs 33.3%, P=0.040), thus improving the intracranial progression-free survival (iPFS) (median iPFS: 11.9 months versus 10.2 months, P=0.039) and overall survival (OS) (median OS: 21.0 months versus 16.7 months, P=0.043). Multivariate analysis showed that WBRT (HR=0.606; 95% CI: 0.403-0.912, P=0.016) and the low Eastern Cooperative Oncology Group performance status (HR=1.884; 95% CI: 1.120-3.170, P=0.017) are independent prognostic factors in all patients. Further subgroup analysis showed that the choice of WBRT time had no effect on patient survival. CONCLUSION: WBRT can improve the survival of patients with multiple BM from NSCLC receiving EGFR-TKI targeted therapy and is an independent prognostic factor. The choice of RT time has no effect on patient survival.

Computed Tomography-guided 125I Radioactive Seed Implantation Therapy for Pancreatic Cancer Pain.

OBJECTIVE: This study aimed to evaluate the short-term curative effect of computed tomography (CT) guided radioactive 125I (125-Iodine) seed implantation (CTRISI) therapy for advanced pancreatic cancer patients pain. STUDY DESIGN: Interventional study. PLACE AND DURATION OF STUDY: Imaging Department and Oncology Department, First Affiliated Hospital of Zhengzhou University, China, from January 2015 to March 2017. METHODOLOGY: A total of thirty-seven patients with advanced pancreatic cancer pain who underwent CTRISI were enrolled. Exclusion criteria were coagulation disorder, multi-organ failure, co-existent pancreatitis and/or uncontrolled hyperglycemia. Cancer pain levels and daily hydroxycodone dose were compared before and after CTRISI, using the Numerical Rating Scale (NRS). Complication after procedure were also noted. RESULTS: All patients were successfully implanted with 125I seeds via CT. After 1-week, 1-month and 2-month of implantation, the NRS scores were significantly lower than preoperative scores (p<0.05). The daily dose of hydroxycodone in one week and one month after operation were significantly lower than preoperation (p<0.001). However, after 2-month of implantation, the daily hydroxycodone dose was not different from the preoperative dose (p=0.198). No patient experienced serious complications. CONCLUSION: CTRISI could relieve pain effectively in the patients and suppress local tumor progress. This technique helps to relieve the pain symptoms, reduce the amount of painkillers, and improve the quality of life of patients. Key Words: Pancreatic cancer, 125I, radioactive seed implantation, pain, hydroxycodone.

Pretreatment NRS-2002 scores combined with hematologic inflammation markers are independent prognostic factors in patients with resectable thoracic esophageal squamous cell carcinoma.

BACKGROUND: The purpose of this study was to investigate the prognostic values of Nutritional Risk Screening 2002 (NRS-2002) and hematologic inflammation markers in patients with esophageal squamous cell carcinoma (ESCC) receiving curative esophagectomy. MATERIALS AND METHODS: A total of 277 patients with ESCC treated with standard curative esophagectomy were retrospectively analyzed. These patients were grouped for further analysis according to the systemic inflammation score (SIS), the combination of neutrophil-to-lymphocyte ratio and platelet-to-lymphocyte ratio (CNP) score and NRS-2002 score. The Kaplan-Meier method and log-rank test were adopted to calculate and compare the progression-free survival (PFS) and overall survival (OS) rates with these parameters. The Cox proportional hazards model was used to carry out univariate and multivariate analyses. Receiver operating characteristic (ROC) curves were applied to verify the accuracy of SIS, CNP and NRS-2002 for survival prediction. RESULTS: In univariate analysis, the following factors were significantly associated with poor PFS and OS: sex, T stage, N stage, TNM stage, SIS, CNP and NRS-2002 (all P<0.05). Furthermore, multivariate Cox regression analysis showed that CNP (hazard ratio [HR]=1.602; 95% confidence interval [CI] 1.341-1.913; P=0.000), NRS-2002 (HR=2.062; 95% CI 1.523-2.792; P=0.000) and TNM stage (HR=1.194; 95% CI 1.058-1.565; P=0.048) were independent prognostic factors for PFS. Correspondingly, CNP (HR=1.707; 95% CI 1.405-2.074; P=0.000), NRS-2002 (HR=2.716; 95% CI 1.972-3.740; P=0.000) and TNM stage (HR=1.363; 95% CI 1.086-1.691; P=0.036) were also independent prognostic factors for OS. Finally, the results of ROC curves indicated that CNP and NRS-2002 were superior to SIS as a predictive factor for PFS or OS in patients with ESCC receiving surgery. CONCLUSION: This study demonstrated that CNP combined with NRS-2002 is promising as a predictive marker for predicting clinical outcomes in patients with ESCC receiving surgery.

Marked Response to Icotinib in Lung Squamous Cell Carcinoma with EGFR Wild-Type.

We present a case of a 72-year male with primary lung squamous cell carcinoma without an epidermal growth factor receptor (EGFR) gene mutation that exhibited a lasting efficacy to icotinib treatment. Originally, the patient was treated with GP regimen for 2 cycles, local lesion radiotherapy (DT46Gy/2Gy*23F), and TP regimen for 2 cycles, with efficacy evaluation as stable disease (SD). The condition was stable for 10 months until the fifth lumbar metastasis was discovered by emission computed tomography (ECT) in July 2015, and the pulmonary metastasis increased. Icotinib was administered at 125 mg orally three times per day. Chest computed tomography (CT) scan showed the tumor was in partial response (PR) and the PR lasted for 27 months.

Celecoxib enhances the radiosensitivity of HCT116 cells in a COX-2 independent manner by up-regulating BCCIP.

It has been reported that celecoxib, a cyclooxygenase-2 (COX-2)-selective nonsteroidal anti-inflammatory drug (NSAID), regulates the radiosensitivity of several cancer cells. BCCIP (BRCA2 and CDKN1A interacting protein) plays a critical role in maintaining the critical functions of p53 in tumor suppression and response to therapy. However, whether the effect of celecoxib on the radiosensitivity of colorectal cancer (CRC) cells is dependent on BCCIP is largely unclear. In this study, we found that celecoxib enhanced the radiosensitivity of HeLa (a human cervical carcinoma cell line), A549 (a human lung carcinoma cell line), and HCT116 cells (a human CRC cells line). Among these cells, COX-2 expression was undetected in HCT116 cells. Treatment with celecoxib significantly increased BCCIP expression in COX-2 negative HCT116 cells. Knockdown of BCCIP obviously abrogated the enhanced radiosensitivity of HCT116 cells induced by celecoxib. A combination of celecoxib and irradiation treatment induced much more γ-H2AX foci formation, higher levels of radiation injury-related proteins phosphorylation, G2/M arrest, apoptosis, and p53 and p21 expression, and lower levels of Cyclin B1 in HCT116 cells than those in cells treated with irradiation alone. However, these changes were undetected in BCCIP-silenced HCT116 cells. Therefore, these data suggest that BCCIP gene may be a radiosensitivity-related gene in CRC. Celecoxib affects the functions of p53 and inhibits the recovery from the irradiation-induced injury by up-regulating the expression of BCCIP, and subsequently regulates the expressions of genes such as p21 and Cyclin B1 to enhance the radiosensitivity of HCT116 cells in a COX-2 independent manner.

An Appreciation for the Rabbit Ladderlike Modeling of Radiation-induced Lung Injury with High-energy X-Ray.

BACKGROUND: To evaluate the utility of rabbit ladderlike model of radiation-induced lung injury (RILI) for the future investigation of computed tomography perfusion. METHODS: A total of 72 New Zealand rabbits were randomly divided into two groups: 36 rabbits in the test group were administered 25 Gy of single fractionated radiation to the whole lung of unilateral lung; 36 rabbits in the control group were sham-radiated. All rabbits were subsequently sacrificed at 1, 6, 12, 24, 48, 72 h, and 1, 2, 4, 8,1 6, 24 weeks after radiation, and then six specimens were extracted from the upper, middle and lower fields of the bilateral lungs. The pathological changes in these specimens were observed with light and electron microscopy; the expression of tumor necrosis factor-α (TNF-a) and transforming growth factor-ß1 (TGF-ß1) in local lung tissue was detected by immunohistochemistry. RESULTS: (1) Radiation-induced lung injury occurred in all rabbits in the test group. (2) Expression of TNF-a and TGF-ß1 at 1 h and 48 h after radiation, demonstrated a statistically significant difference between the test and control groups (each P < 0.05). (3) Evaluation by light microscopy demonstrated statistically significant differences between the two groups in the following parameters (each P < 0.05): thickness of alveolar wall, density of pulmonary interstitium area (1 h after radiation), number of fibroblasts and fibrocytes in interstitium (24 h after radiation). The test group metrics also correlated well with the time of postradiation. (4) Evaluation by electron microscopy demonstrated statistically significant differences in the relative amounts of collagen fibers at various time points postradiation in the test group (P < 0.005), with no significant differences in the control group (P > 0.05). At greater than 48 h postradiation the relative amount of collagen fibers in the test groups significantly differ from the control groups (each P < 0.05), correlating well with the time postradiation (r = 0.99318). CONCLUSIONS: A consistent and reliable rabbit model of RILI can be generated in gradient using 25 Gy of high-energy X-ray, which can simulate the development and evolution of RILI.

High-mobility group boxes mediate cell proliferation and radiosensitivity via retinoblastoma-interaction-dependent and -independent mechanisms.

Our previous studies have shown that high-mobility group box 1 (HMGB1) could physically associate with the retinoblastoma (RB) protein via an LXCXE (leucine-X-cysteine-X-glutamic; X=any amino acid) motif. An identical LXCXE motif is present in the HMGB1-3 protein sequences, whereas a near-consensus LXCXD (leucine-X-cysteine-X-asparagine; X=any amino acid) motif is found in the HMGB4 protein. In this study, we have demonstrated that like HMGB1, HMGB2-3 also associated with the RB in vitro and in vivo, as evidenced by glutathione-s-transferase capture and immunoprecipitation-Western blot assays. A point mutation of the LXCXE or LXCXD motif led to disruption of RB:HMGB1-4 interactions. Enforced expression of HMGB1-3 or HMGB4 by adenoviral-vector-mediated gene transfer resulted in significant inhibition of breast cancer cell proliferation through an LXCXE- or LXCXD-dependent mechanism and an increased radiosensitivity through an LXCXE- or LXCXD-independent mechanism. These results suggest an important role of the LXCXE/D motif in RB:HMGB1-4 association and modulation of cancer cell growth, but not radiosensitivity.

Radio-sensitization of SHG44 glioma cells by Aidi injection in vitro.

In this study, we aimed to explore the radio-sensitization of the SHG44 glioma cell line by Aidi injection and the possible mechanisms involved. The growth curve, cloning efficiency and divisional index of the SHG44 cell line were observed. The inhibition ratio was determined by MTT assay, the change in the cell cycle was analyzed by flow cytometry and the expression of cyclin B1 and Wee1 was detected by western blot analysis. The reproductive activity of the group treated with irradiation (IR) and Aidi injection was suppressed significantly, and the cloning efficiency and divisional index also declined. Aidi injection (15 µg/ml) induced G2/M phase arrest efficiently in the cell line after 48 h. The expression of cyclin B1 decreased in the group treated with IR and Aidi injection compared with either of those with IR or Aidi injection alone. The expression of Wee1 increased in the group treated with IR and Aidi injection compared with that in the groups treated with either IR or Aidi injection alone. In conclusion, Aidi injection is effective in radio-sensitization. The possible mechanisms involved may be associated with G2/M phase cell arrest, the downregulation of cyclin B1 and upregulation of Wee1 expression.

[Influence of knock-down of miR-21 expression on the radiosensitivity of glioma SHG-44 cells].

OBJECTIVE: To explore the influence of knock-down of miR-21 expression on the radiosensitivity of radioresistant glioma SHG-44 cells and its mechanism. METHODS: Radioresistant cell line SHG-44(R) cells were established from radiosensitive parental SHG-44 glioma cells. Then the expression levels of miR-21 in SHG-44(R) and SHG-44 were determined by quantitative real-time PCR. The effect of miR-21 on the radiosensitivity was assessed in SHG-44(R) with miR-21 inhibitor to decrease the miR-21 expression. RESULTS: miR-21 was up-regulated 1.49-fold in SHG-44(R) cells relative to the SHG-44 cells. But after transfection with As-miR-21, the miR-21 expression in SHG-44(R) cells was knocked down significantly. As-miR-21 combined with radiation could synergistically enhanced mitotic death and apoptosis of SHG-44(R) cells, with SER of D(0) and D(q) being equal to 1.48 and 1.54, respectively. Expression levels of caspase-3 in the radiation group, AS-miR-21 transfection group and combination group were 2.24 ± 0.14, 2.05 ± 0.19 and 5.72 ± 0.45, respectively, and its expression in the combination group was higher than that in the other two groups. CONCLUSIONS: miR-21 may be involved in the formation of radioresistance of glioma cells and as a potential target for enhancing the response of radioresistant-glioma cells to radiotheapy.