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Objective:To investigate the effect of heat shock protein 90 (Hsp90) inhibitor PU-H71 combined with X-ray on radioresistant human cervical cancer cells.Methods:The expression levels of Hsp90 gene between cervical cancer tissues and adjacent tissues were analyzed by bioinformatics. Radioresistant cervical cancer cell lines HeLa RR and SiHa RR were obtained by fractional irradiations (2 Gy per fraction, 30 fractions). The cell lines were divided into the control group (treated with dimethyl sulfoxide), irradiation alone group, PU-H71 group (treated with 0.5 μmol/L PU-H71), and PU-H71+irradiation group (irradiation at 24 h after treatment with 0.5 μmol/L PU-H71). Cell survival was detected by clonal formation assay. Immunofluorescence assay was used to detect γH2AX foci at 1, 6, and 24 h after cell treatment. The expression level of Rad51 protein at 1, 2, 6, 12, and 24 h after cell treatment was detected using Western blot. The expression level of phosphorylated DNA-dependent protein kinase catalytic subunit (p-DNA-PKcs) was measured at 2 h after cell treatment. Cell apoptosis at 48 h after cell treatment was assessed by flow cytometry. Results:PU-H71 enhanced the sensitivity of radioresistant cervical cancer cells to X-ray. Compared with the irradiation alone group, the radiation sensitization ratios (SER) of HeLa RR and SiHa RR cells at 10% survival were 1.36 and 1.27, and the apoptosis rates were increased by approximately 72.1% and 63.1% in the PU-H71+irradiation group, respectively. PU-H71 delayed the duration of γH2AX foci induced by X-ray, inhibited the phosphorylation of DNA-dependent protein kinase catalytic subunit (DNA-PKcs), thus preventing non-homologous end joining (NHEJ) repair and delaying homologous recombination repair.Conclusion:PU-H71 increases the radiosensitivity of radioresistant cervical cancer cells by inhibiting the repair pathway of DNA double-strand break, which is expected to be a radiosensitizer to enhance the efficacy of radiotherapy for cervical cancer.
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Objective:To investigate the effect of Salvia miltiorrhiza injection combined with human adipose derived stem cells (hADSC) on the survival rate of autologous fat granules implanted in nude mice.Methods:A total of 24 healthy 8 weeks old female nude mice weighing (23±3) g were randomly divided into four groups ( n=6): Group A was given fat granules 0.5 ml; Group B: fat granules 0.5 ml+ hADSC; Group C: Salvia miltiorrhiza 0.5 g/(kg·d) + fat granules 0.5 ml, and Group D: Salvia miltiorrhiza injection 0.5 g/(kg·d) + fat granules + enrichment. 3 nude mice were randomly selected from each group for 15 days after transplantation and stained with conventional HE. Immunohistochemical staining was performed to count the number of microvessels. On the 30th day after surgery, the remaining 3 nude mice in each group were sacrificed. The specimens were stained with HE and the volume of each specimen was measured. Results:Graft appearance was observed by naked eye: 15 days after the operation, all the specimens were formed completely. The new capillaries were shaped on the surface of the capsule. The boundaries of the capsule and the surrounding tissue were obvious. The activity was good, the hardness of the texture was medium, and the loose connective tissue was connected to the surrounding tissue. On the 30 day after operation, the volume of the graft was smaller than that at the beginning of transplantation, and fat liquefaction and necrosis were seen in some tissues. Blood vessel density values of immunohistochemical sections of fat transplantation in each group 15 days after surgery were compared pairwisely. The differences between the groups were statistically significant ( P<0.05). Lsd-t method was used for pairwise comparison of fat graft volume values of each group 30 days after surgery, and the difference between each group was statistically significant ( P<0.05). Conclusions:The combined use of Salvia miltiorrhiza injection and hADSCs can effectively promote the reconstruction of the early vascular system of the fat granule transplantation and improve the survival rate of fat particles more effectively than the individual use.
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Objective To evaluate the effect of aminolevulinic acid-based photodynamic therapy (ALA-PDT) on the expression of protein kinase D1 (PKD1) in a cutaneous squamous cell carcinoma cell line A431,and to explore the mechanism underlying ALA-PDT-induced apoptosis of A431 ceils.Methods A431 cells were cultured in vitro,and cell counting kit-8 (CCK-8) assay was performed to select the optimal combination of ALA concentration and PDT dose with the strongest proliferation inhibitory effect.A431 ceils at exponential growth phase were randomly divided into 4 groups:control group receiving no treatment,ALA group treated with ALA solution alone,PDT group treated with PDT alone,and ALA-PDT group treated firstly with ALA solution and then with PDT.After 12-,24-,36-and 48-hour additional culture,CCK-8 assay was conducted to evaluate the cellular proliferation inhibition,and the apoptosis rate at the time point of the strongest proliferation inhibitory effect was measured by flow cytometry.RT-PCR was performed to determine the expression of protein kinase D1 gene (PRKD1) in A431 cells at different time points after the ALA-PDT treatment,and Western blot analysis to measure protein expression of PKD 1 and its phosphorylation at Tyr463 (pTyr463) and Ser916 (pSer916) in A431 cells.Results The combination of ALA at the concentration of 1.5 mmol/L with PDT at an irradiation dose of 2 J/cm2 was optimal due to its strongest proliferation inhibitory effect.After 12-,24-,36-and 48-hour additional culture,there were significant differences in the proliferation inhibition rate among the 4 groups (F =39.56,P ＜ 0.05).At 24 hours after the treatment,the ALA-PDT group showed significantly higher proliferation inhibition rate (46.26％ ± 1.25％) compared with the ALA group (14.65％ ± 0.33％,P ＜ 0.05),PDT group (14.96％ ± 0.68％,P ＜ 0.05) and control group (11.98％ ± 0.32％,P ＜ 0.05),as well as compared with that at 12 hours (P ＜ 0.05).At 24 hours after the treatment,the apoptosis rate significantly differed among the 4 groups (F =16.32,P ＜ 0.05),and the ALA-PDT group showed a significantly higher apoptosis rate (41.92％ ± 3.23％) compared with the control group (4.67％ ± 0.88％,P ＜ 0.05),ALA group (7.02％ ± 1.52％,P ＜ 0.05) and PDT group (8.37％ ± 0.59％,P ＜ 0.05).At 0,6,12,24,36 and 48 hours after the treatment,there were significant differences in the mRNA expression of PRKD 1 among the 4 groups (F =22.24,P ＜ 0.05),and the mRNA expression of PRKD1 at 24 hours was significantly lower than that at 0,6,12 hours (all P ＜ 0.05),but was not significantly different from that at 36 and 48 hours (both P ＞ 0.05).No significant difference in the Ser916-phosphorylated PKD1 expression was found among the 4 groups (F =1.53,P ＞ 0.05),while there were significant differences in the expression of PKD1 and Tyr463-phosphorylated PKD 1 among the 4 groups (F =10.04,8.27,both P ＜ 0.05).Additionally,the ALA-PDT group showed significantly lower expression of PKD 1 and Tyr463-phosphorylated PKD 1 compared with the control group,ALA group and PDT group (all P ＜ 0.05).Conclusion PKD1 may be involved in the photochemical process of A431 cell apoptosis induced by ALA-PDT,and may promote the occurrence of squamous cell carcinoma by Tyr463 phosphorylation.
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Objective To measure the expression of protein kinase D1 (PKD1),tyr463-phosphorylaed PKD1 (pPKD1-tyr463) and ser916-phos-phorylaed PKD1 (pPKD1-ser916) in squamous cell carcinoma (SCC),Bowen's disease (BD) and actinic keratosis (AK),and to explore their significance.Methods Fresh tissue samples were resected from lesions of patients with SCC (SCC group),BD (BD group) and AK (AK group),as well as from normal skin of healthy human controls (control group),and each group had a sample size of 10.Real-time RT-PCR was performed to measure the mRNA expression of protein kinase D1 gene (PRKD1),and Western blot analysis to determine the protein expression of PKD1,pPKD1-tyr463 and pPKD1-ser916.In addition,immunohistochemical study was conducted to determine the expression of PKD1,pPKD1-tyr463 and pPKD1-ser916 in another 50 paraffin-embedded skin samples of SCC,20 samples of BD,20 samples of AK and 10 normal skin samples.Results PRKD1 mRNA expression significantly differed among the control group (0.64 ± 0.09),SCC group (5.37 ± 1.06),BD group (2.69 ± 0.72) and AK group (2.43 ± 0.46) (F =21.37,P ＜ 0.05),and was significantly higher in the SCC,BD and AK groups than that in the control group (P ＜ 0.05),as well as in the SCC group than that in the AK and BD groups (both P ＜ 0.05).However,no significant difference in the PRKD1 mRNA expression was observed between the BD group and AK group (P ＞ 0.05).Immunohistochemical study showed that the total PKD1 protein and pPKD1-tyr463 in the SCC and BD groups were mainly expressed in the cytoplasm and cell membrane of spinous layer cells and atypical cells,and their expression rates were significantly higher than those in the AK group and control group (all P ＜ 0.01).The pPKD1-ser916 was only slightly expressed in some cancer nests of well-differentiated SCC tissues,but not in poorly-differentiated SCC,AK,BD tissues and normal skin tissues.In the SCC group,the expression rate of PKD1 increased with the increase of the pathological grade of SCC,and the PKD1 expression was positively correlated with pPKD1-tyr463 expression (rcc =0.479,P ＜ 0.05).Western blot results were consistent with immunohistochemical findings.Conclusion PKD1 and pPKD1-tyr463 may be involved in the development and differentiation of skin tumors derived from stratified squamous epithelium,and PKD1 may exert promotive effects on the formation of cutaneous SCC by activating the Tyr463 phosphorylation site.
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Objective To systematically review the efficacy and safety of postoperative radiotherapy and postoperative chemotherapy for patients with endometrial cancer,which may give support for clinical proper selection.Methods The randomized controlled trials (RCTs)comparing postoperative radiotherapy with post-operative chemotherapy for patients with endometrial cancer were searched in EMBase,PubMed,Cochrane Library,Chinese Biomedical Literature Data,China National Knowledge Infrastructure,and VIP database from the inception to August 201 5.Two reviewers independently assessed the quality of included studies and extrac-ted data.We analyzed the statistic data using RevMan 5.1 software.Results Three RCTs concluding 1 1 21 patients were included.Meta analysis showed that there were no significant differences between the two groups in five-year survival rate (RR =0.94,95%CI:0.80-1 .1 0,Z =0.77,P =0.440),five-year progression-free survival rate (RR =0.98,95%CI:0.90-1 .07,Z =0.52,P =0.61 0)and recurrence rate (RR =1 .06, 95%CI:0.91 -1 .24,Z =0.75,P =0.450),but there were significant differences between the two groups in grade 3-4 thrombocytopenia (RR =0.1 3,95%CI:0.07-0.27,Z =5.62,P <0.000 01 )and grade 3-4 neu-tropenia (RR =0.01 ,95%CI:0.00-0.03,Z =8.27,P <0.000 01 ).Subgroup analysis showed that there were significant differences between the two groups in five-year survival rate (RR =0.79,95%CI:0.68-0.91 , Z =3.1 5,P =0.002)and five-year progression-free survival rate (RR =0.82,95%CI:0.69-0.97,Z =2.31 ,P =0.020)for patients with Ⅲ-Ⅳ stage endometrial cancer.Conclusion Current evidence indicates that compared with postoperative radiotherapy,postoperative chemotherapy may improve the survival rate for pa-tients with advanced stage endometrial cancer.The long-term curative effects still need to be confirmed by RCTs with high quality and large sample.
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OBJECTIVE:To provide optimal therapeutic scheme for patients with acne vulgaris. METHODS: By a single-blind randomized controlled method,a total of 122 patients with acne vulgaris were assigned to one of the following 3 groups: tazarotene cream group(n=42),Water chlorine tincture group(n=38) and combined treatment group(n=42). The curative efficacy was observed after treatment for 8 consecutive weeks. RESULTS: The number of skin lesions in all the 3 groups reduced as the course of treatment increased,and there were significant differences across the 3 groups after 8-week treatment as compared with before treatment (P0.05). CONCLUSION: Tazarotene cream is effective for non-inflammatory acne. The superior efficacy of water chlorine tincture as compared with tazarotene for patients with impetigo might be related to the antibacterial action of chloramphenicol. Tazarotene cream combined with local application of 0.5% water chlorine tincture is effective for inflammatory acne.