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Objective@#To investigate whether cancer-associated- fibroblasts (CAF), the key component of tumor microenvironment, regulate the chemoresistant capacity of lung cancer cell line A549 through SDF-1 secretion.@*Methods@#Primary cell isolation techniques was used to isolate cancer-associated-fibroblasts from lung cancer patients. MTT assay was applied to determine the proliferation and chemoresistance of A549 cells. Quantative PCR was used to detect the mRNA changes of Bcl-xL. Western blotting was used to detect the protein expression of Bcl-xL. ELISA was applied to detect the SDF-1 secretion from normal fibroblasts (NF) and CAF.@*Results@#CAF promoted the proliferation of A549 cells, while NF had no significant effect on them. After 72 hrs incubation, the absorbance value of A549+ CAF medium group was 0.814±0.006, significantly different from the 0.753±0.006 of the A549+ NF medium group (P<0.05). The Q-PCR assay indicated that mRNA expressions of Bcl-xL in the A549 group, A549+ NF medium group and A549+ CAF medium group were 1.00±0.11, 1.10±0.09 and 3.50±0.30, respectively, showing a significant difference between the A549+ NF medium group and A549+ CAF medium group (P<0.05). The Western blot showed that protein expressions of Bcl-xL in the A549 group, A549+ NF medium group and A549+ CAF medium group were 1.00±0.08, 1.10±0.12 and 3.10±0.25, respectively, with a significant difference between the A549+ NF medium group and A549+ CAF medium group (P<0.05). The ELISA results showed that the SDF-1 concentrations in the A549+ NF medium group and A549+ CAF medium group were 3.23±0.02 and 9.53±0.10, respectively, significantly different from each other (P<0.05). The MTT assay indicated that the absorbance values of OD of A549 group, A549+ AMD3100 group, A549+ NF medium group, A549+ NF medium+ AMD3100 group, A549+ CAF medium and A549+ CA Fmedium+ AMD3100 group were 0.43±0.03, 0.25±0.02, 0.48±0.03, 0.31±0.03, 0.72±0.06 and 0.45±0.03, respectively. The data of A549+ NF medium group was significantly different from that of A549+ CAF medium group (P<0.05).@*Conclusions@#Cancer-associated-fibroblasts enhance the drug resistance of A549 cells through SDF-1 secretion, upregulating the expression level of Bcl-xL through interaction with CXCR4. Our study not only illustrates that tumor microenvironment is able to enhance drug resistance of tumor, but also provides experimental evidence for the cancer-associated-fibroblasts as a potential therapeutic target for the treatment of lung cancer.
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Objective To investigate the inhibition mechanism of tetramethylpyrazine combined with cisplatin on angiogenesis in Lewis lung cancer mice and to observe the mechanism of Arresten on angiogenesis in lung cancer. Methods The model of Lewis lung adenocarcinoma mouse xenograft was established in this work, and 40 mice were randomly divided into 4 groups: 0.9% sodium chloride solution group(NS group), tetramethylpyrazine group(TMP group), cisplatin group(DDP group), tetramethylpyrazine plus cisplatin group(TMP + DDP group), 10 mice in each group.Mice in NS group were given 0.2 mL of 0.9% sodium chloride solution, mice in DDP group were given 0.2 mL of 2 mg.kg-1 of cisplatin, mice in TMP group were given 0.2 mL of 100 mg.kg-1 of tetramethylpyrazine, mice in TMP+DDP group were given 2 mg.kg-1 of cisplatin and 100 mg.kg-1 of tetramethylpyrazine, each 0.1 mL .Tumor size was measured every day to calculate the tumor volume.The mice were sacrificed to stripp the subcutaneous tumor after continuous medication. The expressions of Arresten, integrin α1β1 and VEGF were determinated by immunhistochemistry and Western blotting. Results The tumor growth of NS group was the fastest and TMP+DDP group was the slowest. Compared with NS group, the expression of Arresten in the other three groups was increased( P<0.01) , and the TMP+DDP group exhibited the highest expression;at the same time, integrin α1β1 , VEGF in the other three groups was decreased(P<0.01), and the TMP+DDP group exhibited the lowest expression.The expression of integrinα1β1 and VEGF was negatively related to Arresten, and the expression of integrin α1β1 was positively correlated with VEGF. Conclusion TMP can inhibited the growth of Lewis lung carcinoma and angiogenesis. Moreover, in combination with cisplatin, TMP can also improved the effect of chemotherapy and then the survival state of mice. The mechanism of action, which TMP suppress tumor angiogenesis may be through improving Arresten and inhibiting integrin α1β1 and VEGF. And the action mechanism of Arresten may be implemented by inhibiting the expression of VEGF by incorporation with integrinα1β1 or by itself to inhibit the expression of VEGF.
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Objective To investigate the effect of tetramethylpyrazine (TMP) combined with cisplatin(DDP) on the expression of Mac2-binding protein(Mac2-BP) and vascular endothelial growth factor (VEGF) in mice with lung cancer. Methods C57 BL/6 mice were subcutaneously inoculated with Lewis lung adenocarcinoma cells , and Lewis lung adenocarcinoma mouse xenograft model was established .Forty mice were randomly divided into four groups:normal saline group(NS group,0.9%NaCl,0.2 ml), TMP group (TMP 100 mg/kg,0.2 ml), DDP group (DDP 2 mg/kg,0.2 ml), TMP plus DDP group (doses as above,0.2 ml totally) with 10 mice in each group.After 2 drug intervention,following 14 days of inoculation , the tumor diameter was measured every two days to calculate the tumor volume .The mice were sacrificed after 14 days of continuous medication , and the subcutaneous tumors were weighed after stripping to calculate the inhibitory rate.The expressions of Mac2-BP and VEGF was detected by Western blot and immunhistochemistry .Results Compared with NS group, the tumor growth rate of TMP group , DDP group and TMP+DDP group was slowed down , and the tumor growth rate in the TMP+DDP group was decreased most significantly .The inhibitory rate of TMP group , DDP group and TMP+DDP group was 25.57%, 45.24% and 66.5%,respectively.Kim, s formula was used to evaluate the synergy of the combined treatment .The q values for TMP +DDP group was 0.85
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Purpose:To evaluate the efficacy of radiotherapy and chemotherapy in patients with stage Ⅰ and Ⅱ non small cell lung cancer.Methods:From 1991 to 1996,21 patients with stage Ⅰ and Ⅱ non small cell lung cancer were treated.According to the 1997 UICC Staging System, 3 patients had stage Ⅰb disease, 2 stage Ⅱa,and 16 stage Ⅱb.16 patients were confirmed by histopthology,and 5 cases by cytopathology.18 patients had squamous cell carcinoma,2 adenocarcinoma,and 1 mixed squamous cell and adenocarcinoma.21 patients were treated by 6 Mv X or 18 Mv X, received 56 Gy～70 Gy (total dose), at 2 Gy per day.Chemotherapy: 19 patients were treated by MVP(MMC?VDS?DDP). 2 squamous patients were not treated with chemotherapy.19 patients were treated with chemotherapy after radiotherapy 1～7 weeks,and were treated 1～5 cycles.Results:The overall 5 year survial for all patients was 19%.19 patients have died,13 died of tumor,and because of local failure and metastasis.6 patients died of other causes.Conclusions:Non operative treatment is an effective treatment for non small cell lung cancer.