Exosomes derived from hTERT-immortalized cells delay cellular senescence of human fibroblasts.

hTERT gene therapies hold significant promise for treating age-related diseases. However, further research is required to address the challenges of delivery and ethical considerations. We hypothesized that exosomes derived from hTERT-immortalized cells could function similarly to hTERT gene therapies by maintaining telomere length and attenuating cellular senescence biomarkers. In this study, we overexpressed the hTERT gene in Human Foreskin Fibroblast-1 cells (HFF cells) to produce hTERT-immortalized HFF cells (hT-HFF cells). We then used exosomes derived from these hT-HFF cells to treat human fibroblasts, HFF cells. Our results demonstrated that these exosomes effectively attenuated biomarkers of cellular senescence in HFF cells. Furthermore, analysis revealed that hTERT mRNA was indeed packaged into the exosomes from hT-HFF cells. This mRNA was capable of elongating telomeres and delaying cellular senescence in HFF cells. Therefore, exosomes from hT-HFF cells show potential as a treatment for age-related diseases.

EGFR Inhibitor CL-387785 Suppresses the Progression of Lung Adenocarcinoma.

OBJECTIVE: This study aimed to explore the influence of the irreversible EGFR inhibitor CL-387785 on invasion, metastasis, and radiation sensitization of non-small cell lung cancer cells. METHODS: The proliferation inhibitory rate at different time points was detected by MTT assay. The apoptosis of H1975 cells treated with CL-387785 was detected using flow cytometry. The invasion and migration of H1975 cells treated with CL-387785 were determined by Transwell assay and wound healing assay. The survival fraction (SF) of H1975 cells cultured with CL- 387785 under X-ray (0, 2, 4, 6, 8, and 10 Gy) was detected by cloning formation experiment, and the sensitization ratio (SER) was calculated by clicking the multi-target model to fit the cell survival curve. RESULTS: CL-387785 restrained H1975 cell proliferation in a concentration- and time-dependent manner. CL-387785 promoted H1975 cell apoptosis and reduced cell migration distance and the number of transmembrane cells. The SF treated by different concentrations of CL-387785 (10, 25, 50, and 100 nM) was all below 0 nM. The radiation SER of CL-387785 (10, 25, 50 and 100 nM) were 1.17, 1.39, 2.88, and 3.64, respectively. CONCLUSION: The invasion and metastasis of H1975 cells were restrained by irreversible EGFR inhibitor CL-387785. CL-387785 also exhibited the effect of radiotherapy sensitization.

The mechanism of radiotherapy for lung adenocarcinoma in promoting protein SIRT6-mediated deacetylation of RBBP8 to enhance the sensitivity of targeted therapy.

BACKGROUND: Lung cancer has the fastest increase in morbidity and mortality, and is one of the most threatening malignant tumors to human health and life. Both radiotherapy and targeted therapy are typical treatments after lung cancer surgery. Radiotherapy is a means of locally killing cancer lesions, and it plays an important role in the entire management of lung cancer. Gefitinib is one of the most commonly used targeted therapy drugs in the treatment of lung cancer. The purpose of this project is to explore the mechanism by which deacetylation of RBBP8 mediated by radiotherapy-promoting protein SIRT6 in lung adenocarcinoma enhances the sensitivity of targeted therapy. METHODS: In both the cell experiments and the animal experiments, the samples were divided into five groups: Model group, RT group, CT group, RT+CT group, and RT+CT+inhibitor group. The CCK8 method was used to detect the viability of each group of cells. The flow cytometry experiment was used to analyze the apoptotic characteristics of each group of cells. The scratch test was used to detect the migration ability of each group of cells. Transwell invasion test was used to determine the invasion ability of each group of cells. The lung tumor tissues of each group of mice were collected to analyze the tumor size, volume, and metastasis characteristics. The TUNEL experiment was used to detect the apoptosis characteristics of the cells in the lung cancer tissues of each group mice. Immunohistochemistry experiments were used to analyze the distribution and relative expression characteristics of protein SIRT6 in mouse lung cancer tissues. The colorimetric experiments were used to detect the activity of Caspase 3 and Caspase 8 in each group. Western blot method was used to detect the expression of SIRT6, RBBP8, and MYC in each group. RESULTS: In each experiment, the results of the experiment have mutually proven consistency, and there is no contradiction. In addition to the Model group, the other 4 groups used different treatment methods. The better the curative effect, the lower the cell viability of cancer cells and the higher the apoptotic ratio. This is reflected in the CCK8 test, flow cytometry analysis, cell scratch test, Transwell cell migration test, and TUNEL detection. At the same time, colorimetric detection and Western blot analysis also analyzed the levels of SIRT6, RBBP8 and other cancer-related proteins in each group at the molecular level, implying the importance of SIRT6 protein in the treatment process. CONCLUSION: Our project has proved that radiotherapy can promote the protein SIRT6 to deacetylate RBBP8 proteins, and ultimately enhance targeted therapy drug sensitivity.

Hypoxia-induced PVT1 promotes lung cancer chemoresistance to cisplatin by autophagy via PVT1/miR-140-3p/ATG5 axis.

Lung cancer is one of the most common and lethal malignant tumors and the cases increased rapidly. Elevated chemoresistance during chemotherapy resistance remains a challenge. Hypoxia is one of the components that lead to chemoresistance. PVT1 participates in various tumor drug resistance and is associated with hypoxia conditions. The present study aimed to analyze the regulatory relationship of hypoxia and PVT1 and the mechanism of PVT1 in the hypoxia-induced chemoresistance process of lung cancer. The expression of PVT1 in lung cancer and adjacent tissues, and cell lines were analyzed using the TCGA database and qPCR. The regulatory relationship between hypoxia and PVT1 was validated and analyzed with qPCR, luciferase reporter system, and CHIP-qPCR. The role of PVT1 in chemoresistance ability induced by hypoxia was analyzed with CCK-8 assay and flow cytometry. The roles of PVT1, hypoxia, and chemoresistance were also analyzed with LC3-GFP transfection, WB, and IHC. Finally, the results were further validated in xenograft models. PVT1 is highly expressed in lung cancer and cell lines, and the expression of PVT1 is regulated by HIF-1α, and the luciferase reporter assay and CHIP-qPCR analysis indicated that HIF-1α could bind to the promoter region of PVT1 and regulate PVT1 expression. PVT1 participated in hypoxia-induced chemoresistance and induced higher viability and lower apoptosis rate by the autophagy signaling pathway via PVT1/miR-140-3p/ATG5 axis. All the findings were validated in the xenograft models. In conclusion, these results suggest that the expression of PVT1 is regulated by HIF-1α and participates in hypoxia-induced chemoresistance.

Xanthorrhizol inhibits non-small cell carcinoma (A549) cell growth and promotes apoptosis through modulation of PI3K/AKT and NF-κB signaling pathway.

Xanthorrhizol (XNT) is a sesquiterpenoid agent isolated from Curcuma xanthorrhiza; It is known to exhibit various pharmacological activities including anti-cancer. We investigated the anti-cell proliferative and proapoptotic effects of XNT on Non-small cell carcinoma (A549) cells were analyzed by the generation of reactive oxygen species (ROS), alteration of mitochondrial membrane potential (ΔΨm), oxidative DNA damage, and apoptosis morphological changes were explored by Hoechst and AO/EtBr staining. Our study demonstrated that XNT treatment significantly reduced the viability of A549 cells in a concentration-dependent manner. We observed that XNT-induced oxidative stress-mediated apoptotic cell death by increasing intracellular ROS generation, depleting antioxidant levels, enhancing lipid peroxidation, increased apoptotic morphological changes, and % of DNA damage on human lung cancer cells. Furthermore, we observed that the XNT induce apoptosis through inhibits phosphorylation of PI3K, AKTand inhibit NF-κBp65 transcriptional signaling activity. In addition, XNT treatment alters the ΔΨm, thereby induces apoptosis was closely coordinated with the induction of pro-apoptotic markers Bax, Bad, caspase- 3, 9 and cytochrome c, and suppression of anti-apoptotic (Bcl-2, Bcl-XL) protein expression. According to our results, XNT-inducing apoptosis in A549 cells by causing oxidative damage and modulating apoptotic signaling events. Finally, XNT-induced apoptotic cell death was confirmed by the TUNEL assay. Therefore, XNT might be used as a chemotherapeutic agent for the treatment of lung cancer.

Experimental studies on the protective effects of the overexpression of lentivirus-mediated sirtuin 6 on radiation-induced lung injury.

BACKGROUND: Sirtuin 6 (SIRT6) can increase the radiosensitivity of non-small cell lung cancer and exert protective effects on radiation-induced lung injury. OBJECTIVES: To investigate protective effects of SIRT6 overexpression on radiation-induced lung injury in rats. MATERIAL AND METHODS: Male Wistar rats (n = 72) were randomly divided into 3 groups. Models were made by radiating both lungs with a 6MV X linear accelerator. Each group was injected through the tail vein with normal saline (the control group and radiation group) and lentivirus carrying overexpressed SIRT6 (the Lent-SIRT6 group) on the same day as the modeling. Routine blood indexes (white blood cells (WBC), red blood cells (RBC), neutrophils and lymphocytes) were recorded; the rats were sacrificed and their lung tissues taken; pathological changes in the lungs were evaluated using hematoxylin and eosin (H&E) staining; and tumor necrosis factor α (TNF-α), interleukin 6 (IL-6) and interleukin 1ß (IL-1ß) were detected with enzyme-linked immunosorbent assay (ELISA) 8 weeks after radiotherapy. RESULTS: The lung structure including alveolar walls and interstitium in the control group was normal, but the alveolar walls in the radiation group were obviously thickened and a large amount of hyperplastic fibrous tissue was found in the alveolar interstitium. The thickness and interstitial fibrosis of the alveolar walls were more alleviated in the Lent-SIRT6 group than in the radiation group. Compared with those in the control group, the respiratory rates, levels of TNF-α and IL-6 in serum, neutrophils and levels of TNF-α, IL-6 and IL-1ß in the liver all were increased, while WBCs and lymphocytes were decreased in the radiation group. The respiratory rates, levels of TNF-α and IL-6 in serum, neutrophils and levels of TNF-α, IL-6 and IL-1ß in the liver were all decreased, and WBCs and lymphocytes were increased after injection with lentivirus carrying overexpressed SIRT6. CONCLUSIONS: Sirtuin 6 inhibits inflammation and alleviates radioactive pneumonia and lung injury. Therefore, SIRT6 can exert certain protective effects on lung injury.

Radiosensitization effects of curcumin plus cisplatin on non-small cell lung cancer A549 cells.

The aim of the present study was to determine the radiosensitization effect of the combination of curcumin and cisplatin on non-small cell lung cancer (NSCLC) A549 cells. Cell viability was analyzed using the MTT assay following treatment with different concentrations of curcumin and cisplatin for 24~72 h. Survival fraction (SF) value of the treatment groups (single irradiation, curcumin + irradiation, cisplatin + irradiation, and curcumin + cisplatin + irradiation) treated with different doses of X-ray radiation were evaluated using colony formation assay, according to a multi-target single-hit model. Migration and invasion as well as the levels of epidermal growth factor receptor (EGFR) protein following 24 h were detected by scratch wound assay, Matrigel assay and western blot analysis, respectively. The results of the present study demonstrated that the viability of the cells decreased after being treated by curcumin, and the inhibitory effect was dose and time-dependent as the concentration of curcumin increased from 10 to 200 µmol/l (P<0.05). SF value was lower in the curcumin + cisplatin + irradiation group compared with the other three treatment groups at 2~10 Gy. Furthermore, SF value was lower in the curcumin + irradiation group at 4~10 Gy. The SF value was also lower in the cisplatin + irradiation group at 2~10 Gy compared with the single irradiation group (P<0.05). The sensitization enhancement ratios in the curcumin + irradiation, cisplatin + irradiation, and curcumin + cisplatin + irradiation groups were 1.24, 1.31 and 1.96, respectively. The migration distance, the number of cells invaded through the transmembrane, and the level of EGFR protein in four treatment groups were the highest in the single irradiation group, compared with the other three treatment groups (P<0.05). Furthermore, the radiosensitization effects of curcumin and cisplatin on NSCLC A549 cells, which include inhibition of proliferation, migration and invasion, may be associated with the inhibition of the EGFR-associated signaling pathway.

LncRNA HMMR-AS1 promotes proliferation and metastasis of lung adenocarcinoma by regulating MiR-138/sirt6 axis.

PURPOSE: Long noncoding RNAs (lncRNA) play critical roles in cancer development. In this study, we aimed to explore the function and possible molecular mechanism of HMMR-AS1 involved in lung adenocarcinoma (LUAD). EXPERIMENTAL DESIGN: Firstly, we analyzed HMMR-AS1 expression in LUAD tissues with the sequencing data from The Cancer Genome Atlas (TCGA). Next, we evaluated the effects of HMMR-AS1 on LUAD cell proliferation and apoptosis, and its regulation of miR-138 by acting as a ceRNA. The animal model was used to support the in vitro experimental findings. RESULTS: HMMR-AS1 expression was significantly upregulated in LUAD tissues and was associated with larger tumor diameter, advanced TNM stage, lymph node metastasis, and shorter survival. Knockdown of HMMR-AS1 induced apoptosis and growth arrest in vitro and inhibited tumorigenesis in mouse xenografts. Mechanistically, HMMR-AS1 functioned as a ceRNA of miR-138, thereby leading to repression of its endogenous target sirt6. Moreover, knockdown of HMMR-AS1 dramatically inhibited tumor growth and metastasis of LUAD in vivo. CONCLUSIONS: Taken together, HMMR-AS1 is significantly over-expressed in LUAD, and HMMR-AS1-miR-138-sirt6 axis play a critical role in LUAD tumorigenesis. Our findings highlight an oncogenic role of HMMR-AS1 in LUAD.

Effects of microRNA-513b on cell proliferation, apoptosis, invasion, and migration by targeting HMGB3 through regulation of mTOR signaling pathway in non-small-cell lung cancer.

This study aimed to explore the underlying mechanism of miR-513b and HMGB3 in regulating non-small-cell lung cancer (NSCLC). NSCLC tumor, adjacent tissues, and cell lines were extracted, and the expression of miR-513b and HMGB3 were determined by quantitative real-time polymerase chain reaction (RT-qPCR) and western blot analysis. Then, miR-513b was overexpressed in NSCLC cell, and the proliferation, migration, invasion, and apoptosis of cells were determined by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT), wound healing, transwell, and flow cytometry, respectively. Regulatory relationship between miR-513b and HMGB3 was determined using luciferase activity reporter assay. Lastly, HMGB3 and/or miR-513b were overexpressed in NSCLC cells, and the proliferation, migration, invasion, and apoptosis of cells were determined. Compared with the controls, the expression of miR-513b was significantly downregulated in the NSCLC tissues and cells lines by RT-qPCR ( p < 0.05). However, the expression of HMGB3 was significantly downregulated at both messenger RNA and protein levels ( p < 0.05). Overexpression of miR-513b could significantly inhibit the proliferation, invasion, migration, and promote apoptosis of NSCLC cells ( p < 0.05). HMGB3 was a target of miR-513b, and overexpression of HMGB3 could obviously reverse the effect of miR-513 on the proliferation, invasion, migration, and apoptosis of NSCLC cells ( p < 0.05). The present results could suggest miR-513b was downregulated in NSCLC and could regulate the proliferation, invasion, migration, and apoptosis of NSCLC cells via HMGB3.

Influence of SIRT6 regulation of cellular glycometabolism on radiosensitivity of non-small-cell lung cancer A549 cells.

OBJECTIVE: To investigate the influence of SIRT6 mediated regulation of cellular glycometabolism on radiosensitivity of A549 non-small-cell lung cancer (NSCLC) cells. METHODS: Ad-SIRT6 adenovirus vector overexpressed SIRT6 and was established and divided into a control group, a zero-load group (Ad-null), and an overexpression group (Ad-SIRT6). The virus concentration of the Ad-null group and the Ad-SIRT6 group was 200 pfu/cell. The survival factor (SF) after X-ray irradiation of 0, 2, 4, 6, 8, and 10 Gy in three groups was detected by clone formation and cell cycle and apoptosis after 4 Gy X-ray irradiation for 48 hours in the three groups was detected by flow cytometry. Expression levels of pyruvate kinase (PKM), lactate dehydrogenase (LDHA), and hexokinase (HK) after 4 Gy X-ray irradiation of 48 h were detected by qPCR. The glucose level after consumption of (1×106) cells in the medium was detected by a glucose kit. RESULTS: Compared with the control group and the Ad-null group, SFs after X-ray irradiation of 4-10 Gy in the Ad-SIRT6 group were decreased (P<0.05). A sensitization enhancement ratio of the Ad-SIRT group/the control group was 1.451. After 4 Gy X-ray irradiation of 48 h, the cell ratio and apoptosis rate in G1 phase were increased in the Ad-SIRT6 group, with statistical significance when compared with the other two groups (P<0.05). Compared with the control group and the Ad-null group, levels of PKM, LDHA, and HK mRNA in Ad-SIRT6 group were decreased (P<0.05) and the remaining glucose in the medium was increased (P<0.05). CONCLUSION: Overexpression of SIRT6 can inhibit key-enzyme generation in A549 cells to inhibit glycolysis, enhance the radiosensitivity, and lead to G0/G1 phase block as well as cell apoptosis.

SIRT6 overexpression inhibits HIF1α expression and its impact on tumor angiogenesis in lung cancer.

OBJECTIVE: To investigate the effect of silencing information regulator 6 (SIRT6) on HIF1α expression of cell line A549 in non-small cell lung cancer and on tumor angiogenesis in lung cancer. METHODS: Cell line A549 in the logarithmic growth phase was transfected with Ad-SIRT6 and Ad-null respectively. According to the study design, the cells were divided into control group, Ad-null group and Ad-SIRT6 group. The HIF1α and HIF2α mRNA expression in each group were detected by real-time quantitative PCR (qPCR). The level of prolyl hydroxylase (PHD) 1-3 after 48 h of Ad-SIRT6-transfected cell line A549 and the levels of VEGF-C, VEGF-D, VEGFR-2 and VEGFR-3 in the supernatants were determined by ELISA. The nude mice were injected subcutaneously with Ad-null or Ad-SIRT6 transfected cell line A549. The tumor volume was observed at 6, 12, 18, 24 and 30 d after inoculation, and the tumor mass was weighed at 30 d. Also, microvessel density (MVD) and the number of positive HIF1α and VEGF cells were detected by immunohistochemistry. The VEGF and HIF1α levels in tumor tissue were detected by ELISA and qPCR respectively. RESULTS: qPCR showed that the levels of HIF-1α mRNA, VEGF-C, VEGF-D, VEGFR-2 and VEGFR-3 in the supernatant were decreased, the level of PHD2 was increased (P<0.05), and the levels of HIF-2α mRNA, PHD1 and PHD3 did not change much (P>0.05) in the Ad-SIRT6 group as compared with those in the control group and Ad-null group. The tumor growth rate was decreased, and the tumor volume at 12-30 d after inoculation was less in the Ad-SIRT6 group than in the control group and Ad-null group (P<0.05); the tumor mass was also lower than that of control and Ad-null groups (P<0.05). Immunohistochemistry showed that MVD and the number of HIF-1α and VEGF positive cells were less in the Ad-SIRT6 group than in control and Ad-null groups (P<0.05); and HIF-1α and VEGF levels in tumor tissue were decreased in the Ad-SIRT6 group compared to the control and Ad-null groups (P<0.05). There were no significant differences in the above measurements between the control group and Ad-null group (P>0.05). CONCLUSION: SIRT6 overexpression can inhibit HIF1α and VEGF expression, promoting PHD2 expression, which can inhibit angiogenesis and xenograft growth and may play a role in reducing HIF1α and VEGF expression.

Elevated Serum Concentration of Chitinase 3-Like 1 is an Independent Prognostic Biomarker for Poor Survival in Lung Cancer Patients.

BACKGROUND/AIMS: The chitinase 3-like 1 (CHI3L1) has an important role in cancer progression, and high CHI3L1 expression is associated with the development and progression of cancers. Previous studies had been controversial with respect to the association between CHI3L1 expression and lung cancer prognosis. Thus, we performed a meta-analysis to investigate the prognostic value of CHI3L1 expression in lung cancer. METHODS: We searched Pubmed, Embase, and Wanfang databases to identify eligible studies. Overall survival and disease free survival were collected from included studies. Pooled hazard ratios (HRs) with 95% confidence interval (95%CI) were calculated to estimate the association. Seven studies comprising 911 lung cancer patients were included in this meta-analysis. RESULTS: The results showed high CHI3L1 expression was independently associated with poorer overall survival in lung cancer patients (HR = 1.71, 95%CI 1.24-2.37, P = 0.001). Subgroup analysis by histological type showed that high CHI3L1 expression was independently associated with poorer overall survival in both non small-cell lung cancer patients (HR = 2.23,95%CI 1.43-3.47, P < 0.001) and small-cell lung cancer patients (HR = 1.45, 95%CI 1.06-2.00, P = 0.021). In addition, sensitivity analysis by omitting single study by turns did not change the pooled outcomes obviously. CONCLUSION: Our results suggest that elevated serum CHI3L1 concentration is an independent prognostic biomarker for poorer survival in lung cancer patients.

Radiosensitization effect of overexpression of adenovirus-mediated SIRT6 on A549 non-small cell lung cancer cells.

OBJECTIVE: To explore the radiosensitization effect of overexpression of silent information regulator 6 (SIRT6) on A549 non-small cell lung cancer (NSCLC) cells. METHODS: Adenovirus vector Ad-SIRT6 causing overexpression of SIRT6 was established. Western blotting and MTT assay were adopted to detect the level of SIRT6 protein and the inhibitory rate of A549 cell proliferation after different concentrations of adenovirus transduction (0, 25, 100, 200, and 400 pfu/cell) for 24 h. Control group, Ad-null group and Ad-SIRT6 group were designed in this experiment and virus concentration of the latter two groups was 200 pfu/cell. Colony formation assays were employed to test survival fraction (SF) of the 3 groups after 0, 2, 4, 6, 8, 10 X-ray irradiation. Flow cytometry was used to detect the status of cell cycle of 3 groups after 48 h of 4 Gy X-ray irradiation and Western blotting was used to determine the expression of apoptosis-related genes of 3 groups after 48 h of 4 Gy X-ray irradiation. RESULTS: In the range of 25 ~ 400 pfu/cell, the inhibitory rate of A549 cell proliferation increased as adenovirus concentration raised. The inhibitory rates under the concentrations of 0, 25, 100, 200, and 400 pfu/cell were 0%, 4.23 ± 0.34%, 12.7 ± 2.57%, 22.6 ± 3.38%, 32.2 ± 3.22%, 38.7 ± 4.09% and 47.8 ± 5.58% and there were significantly differences among groups (P < .05). SF in Ad-SIRT6 group was lower than Ad-null and control groups after 4 ~ 10 Gy X-ray irradiation (P < 0.05) and the sensitization enhancement ratio (SER) was 1.35 when compared with control group. Moreover, after 48 h of 4 Gy X-ray irradiation, there appeared a significant increase in G1-phase cell proportion, up-regulated expression of the level of apoptosis-promoting genes (Bax and Cleaved caspase-3), but a obvious decline in S-phase and G2-phase cell proportion and a significant decrease of the level of apoptosis- inhibiting gene (Bal-2) in the Ad-SIRT6 group (P<0.05). CONCLUSION: The over-expression of adenovirus-mediated SIRT6, which has radiosensitization effect on A549 cells of NSCLC, can inhibit the proliferation of A549 cells and cause G0/G1 phase retardation as well as induce apoptosis of cells.

Effect of Withaferin A on A549 cellular proliferation and apoptosis in non-small cell lung cancer.

OBJECTIVE: To explore the effect of Withaferin A on A549 cellular proliferation and apoptosis in non-small cell lung cancer (NSCLC). MATERIALS AND METHODS: NSCNC cell line A549 was selected to explore the effect of Withaferin A on A549 cellular proliferation, apoptosis and the PI3K/Akt signal pathway capable of regulating tumor biological behavior by assessment of cellular proliferation, cellular apoptotic rates and cellular cycling as well as by immuno-blotting. RESULTS: Withaferin A could inhibit A549 cellular proliferation and the control rate was dosage-dependent (P<0.05), which also increased time-dependently with the same dosage of Withaferin A (P<0.05). The apoptotic indexes in A549 cells treated with 0, 2.5, 5.0, 10.0 and 20.0 µmol·L-1 Withaferin A for 48 h were significantly different (P<0.05). In addition, the apoptotic rates of each group in both early and advanced stages were higher than those in 0 µmol·L-1 (P<0.05), which were evidently higher after 48 h than those after 24 h (P<0.05). A549 cells treated by Withaferin A for 48 h were markedly lower in Bcl-2 level and obviously higher in Bax and cleaved caspase-3 levels than those treated by 0 µmol·L-1 Withaferin A (P<0.05), and there were significant differences among 5, 10 and 20 µmol·L-1 Withaferin A (P<0.05). The ratios of A549 cells treated by Withaferin A for 48 h in G0/G1 stage were higher than those in 0 µmol·L-1 , while those in S and G2/M stages were obviously lower than those in G2/M stage, and there were significant differences in 5.0, 10.0 and 20.0 µmol·L-1 Withaferin A (P<0.05). Additionally, p-Akt/Akt values were in reverse association with dosage, and the differences were significant (P<0.05). CONCLUSION: Withaferin A can inhibit the proliferation and apoptosis of A549 cells by suppressing activation of the PI3K/Akt pathways.

[Analysis of prognostic factors in NSCLC patients with brain metastases 
diagnosed by constrast-enhanced MRI after whole brain radiotherapy].

BACKGROUND AND OBJECTIVE: Brain metastases are a common complication of lung cancer, occurring in 20%-40% of patients. The aim of this study is to explore prognostic factors in non-small cell lung cancer (NSCLC) in patients with brain metastases diagnosed by constrast-enhanced MRI after whole brain radiotherapy. METHODS: TA retrospective review of clinical data from 241 NSCLC patients with brain metastases received whole brain radiotherapy from April 2007 to October 2008 was performed. A number of potential factors that might affect prognosis after irradiation were evaluated. The significance of prognostic variables in the survival resulted from univariate analysis by Kaplan-Meier combining with Log-rank test, and the multivariate analysis was obtained by Cox regression model. RESULTS: Median follow-up time for the survivors was 19.1 months. For all patients, the median survival time (MST) was 8.7 months. By univariate analysis, female patients with KPS>70, no symptom when diagnosed with brain metastases, tumor controlled in the chest, and received more than 3 cycles of chemotherapy and combined target therapy were the important factors for overall survival. By multivariate analysis, female, tumors controlled in the chest, and combined target therapy were independent prognostic factors for NSCLC patients with brain metastases. Tumor controlled in the chest was the most important independent prognostic factor. CONCLUSIONS: Gender, local tumor controlled, and combined target therapy significantly influenced NSCLC brain metastases diagnosed by constrast-enchanced MRI survival after whole brain radiotherapy.eciated patient subsets and is a useful method for dissecting complex clinical situations. Moreover, CART can be used to identify homogeneous patient populations in clinical practice and future clinical trials.

[Prognostic analysis of advanced non small cell lung cancer treated by sequential chemo-radiation therapy combined with traditional Chinese medicine: a report of 54 cases].

OBJECTIVE: To evaluate the influence of fewer courses and prolonged intervals of chemotherapy on survival rate of advanced non small cell lung cancer (NSCLC) patients treated by sequential chemo-radiation therapy combined with traditional Chinese medicine (TCM). METHODS: From Jan. 2000 to Dec. 2001, 54 untreated advanced NSCLC patients (2 stage IIIa, 18 stage IIIb, 34 stage IV) were treated by sequential chemo-radiation therapy combined with TCM. The courses of chemotherapy were reduced and the intervals of chemotherapy were longer than that of the standard regimen. The efficacy and survival rate were documented and the prognostic factors were analyzed. RESULTS: Complete remission (CR) was observed in 1 case and partial remission (PR) in 20 cases. The overall objective response rate was 40.4%. Median survival was 15.3 months, 1-, 2- and 3-year survival rate were 53.7%, 28.9% and 9.6% respectively. The median survival of stage III and IV were 21.8 months and 12.5 months respectively. The 1-, 2-, and 3-year survival rates of stage III were 65.0%, 49.5%, 24.7% and that of stage IV were 47.0%, 23.3%, 0%, respectively. The quality of life was improved in most of the patients. Cox's proportional hazards regression showed that improved quality of life and treatment of TCM were the significant prognostic factors of overall survival. CONCLUSION: Chemotherapy and radiotherapy combined with TCM is beneficial to extending the interval of chemotherapy, improving the quality of life, and increasing the survival rate of advanced NSCLC patients.