1.Expression of stathmin and its influence on taxol sensitivity in esophageal squamous cell carcinoma
Gaijing HAN ; Lu YAN ; Fangfei NIU ; Fang LIU ; Lanping ZHOU ; Xiaohang ZHAO ; Yang XU
Journal of Medical Postgraduates 2016;29(7):677-682
Objective Stathmin, a microtubule-destabilizing protein , has high expression in esophageal squamous cell carci-noma(ESCC), while taxol is a common chemotherapy microtubule-targeted drug for esophageal cancer .This study aimed to investigate the impact of stathmin expression and its influence on taxol sensitivity in ESCC . Methods We established 2 cell models with ST-MN1 gene overexpression in KYSE 510 and KYSE 170 cell lines, including KYSE 510-Stathmin, KYSE 170-Stathmin, KYSE 510-Control and KYSE 170-Control.MTT assay and colony formation were applied to compare the taxol sensitivity between experimental group and control group .Flow cytometry was used to measure the apoptosis of KYSE 510-Stathmin and KYSE 510-Control after taxol treatment.Western blot was used to test the changes of related factors to apoptosis and autophagy . Results ①Stathmin protein ex-pressions in KYSE 510-Stathmin and KYSE 170-Stathmin cells were higher than those of control cells (P<0.01).② The percentages of inhibition were significantly decreased in KYSE 510-Stathmin and KYSE 170-Stathmin cells 24 h after 50, 100,250 nmol/L taxol treat-ment compared with KYSE 510-Stathmin cells(P <0.01).③The percentages of inhibition were significantly reduced in KYSE 510-Stathmin and KYSE 170-Stathmin cells after 250 nM taxol treatment for 24, 48, 60 h (P<0.01).④After taxol treatment,the number of colony formation in KYSE 510-Stathmin cells was higher com-pared with KYSE 510-Control cells (P<0.01).⑤The percentage of cell apoptosis in KYSE 510-Stathmin was significantly lower than that of KYSE 510-Control cells by flow cytometry (11.90%±0.78%vs 29.63%±3.26%, P<0.05).Western blot showed the ap-optosis of associated proteins such as the activation of Caspase 8 and Caspas9. Conclusion The result indicates that overexpression of stathmin inhibits taxol sensitivity in ESCC cell lines .
2. Effect of Beclin1 overexpression on biological behaviors of SK-MEL-2 human malignant melanoma cells
Yanyan XU ; JiuJiang WANG ; Ling ZHANG ; Zhao HAN ; Liang NIU ; Jianzhong ZHANG ; Zhao LIU ; Gaijing LI ; Xiaobing LI ; Qing LIU ; Zhijun LIU ; Xiaojing LI
Chinese Journal of Dermatology 2020;53(1):40-44
Objective:
To evaluate the effect of overexpression of the autophagy marker gene Beclin1 on biological behaviors of SK-MEL-2 human malignant melanoma cells.
Methods:
Western blot analysis was performed to determine the protein expression of Beclin1 in melanoma cell lines A375 and SK-MEL-2. SK-MEL-2 cells with low Beclin1 protein expression were selected as research objects, and divided into 3 groups: blank group receiving no treatment, negative control group transfected with pcDNA.3.1/myc-His (-) A, and experimental group transfected with pcDNA3.1-Beclin1 plasmid. After 2-week culture, cell counting kit-8 (CCK-8) assay was conducted to evaluate the effect of Beclin1 on cell proliferation at 24, 48 and 72 hours, and Transwell assay and wound-healing assay were performed to assess the effect of Beclin1 overexpression on the invasion and migration abilities of SK-MEL-2 cells. Repeated measures analysis of variance and completely randomized analysis of variance were used to analyze differences in indices among groups, and least significant difference (LSD) -