Chinese Medical Sciences Journal ›› 2019, Vol. 34 ›› Issue (3): 184-193.doi: 10.24920/003558
• Original Article • Previous Articles Next Articles
Downregulation of iASPP Expression Suppresses Proliferation, Invasion and Increases Chemosensitivity to Paclitaxel of Head and Neck Squamous Cell Carcinoma In Vitro
Liu Zhengzheng1, *(
), Kuang Weilu1, Zeng Wenjing3, Xiao Jianyun2, Tian Yongquan2, *()
- 1 Department of Oncology, , Xiangya Hospital, Central South University, Changsha 410008, China
2 Department of Pharmacy, , Xiangya Hospital, Central South University, Changsha 410008, China
3 Department of Otolaryngology Head and Neck Surgery, Xiangya Hospital, Central South University, Changsha 410008, China
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Received:2019-07-02Published:2019-09-30Online:2019-07-02 -
Contact:Liu Zhengzheng,Tian Yongquan E-mail:lzzsmile@aliyun.com;tianyongquan@aliyun.com
Cite this article
Liu Zhengzheng, Kuang Weilu, Zeng Wenjing, Xiao Jianyun, Tian Yongquan. Downregulation of iASPP Expression Suppresses Proliferation, Invasion and Increases Chemosensitivity to Paclitaxel of Head and Neck Squamous Cell Carcinoma In Vitro[J].Chinese Medical Sciences Journal, 2019, 34(3): 184-193.
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Figure 1.
Knockdown of iASPP expression by lentiviral vector-mediated shRNA interference in Tu686 cell lines. CON: non-infected cells; shRNA-NC: cells transfected with control shRNA lentiviral vector; shRNA-iASPP: cells transfected with iASPP lentiviral vector. A. Relative iASPP mRNA expression was detected by qPCR. GAPDH was used as an internal control (*P = 0.000, compared with the shRNA-NC group, Student’s t-test, n=3). B. Decreased protein expression of iASPP determined by Western blotting. C. Relative expression levels of iASPP protein (*P = 0.000, compared with the shRNA-NC group, Student’s t-test, n=3)."
Figure 1.
Figure 2.
The effect of iASPP knockdown on the proliferation of Tu686 cells. Cell proliferation was measured by the CCK-8 assay every 24 hour for 5 days. Results were expressed as means of three independent experiments ± SD (*P < 0.01, compared with shRNA-NC cells and CON cells)."
Figure 2.
Figure 3.
The effect of knockdown of iASPP on cell apoptosis of Tu686 cells was analyzed by flow cytometry. Representative histograms showing apoptosis of CON (A), shRNA-NC (B) and shRNA-iASPP cells (C). The apoptosis ratio of shRNA-iASPP cell line was significantly higher than that of CON cell line and shRNA-NC cell line (D, *P = 0.000). Results of quantitative analysis represent the mean ± SD of three independent experiments."
Figure 3.
Figure 4.
The effect of knockdown of iASPP on cell cycle distribution of Tu686 cells was analyzed by flow cytometry. Representative histograms showing cell cycle distribution of CON (A), shRNA-NC (B) and shRNA-iASPP cells (C). The percentage of shRNA-iASPP cells in G0/G1 phase was strikingly increased (*P=0.000), whereas the percentage of cells in S phase and G2/M phase decreased significantly (D) (both *P=0.000). Results of quantitative analysis represent the mean ± SD of three independent experiments."
Figure 4.
Figure 5.
The effect of knockdown of iASPP expression on the expression of apoptosis-related proteins in Tu686 cells was analyzed by western blotting. As shown above, downregulation of iASPP expression obviously inhibited the expression of anti-apoptotic protein bcl-2, but significantly increased pro-apoptotic protein bax expression."
Figure 5.
Figure 6.
Downregulation of the expression of iASPP inhibited the invasion of Tu686 in vitro. Matrigel invasiveness assay in transwell culture chamber showed invasiveness decreased significantly in the shRNA-iASPP group (C), compared with the CON group (A) and shRNA-NC group (B)."
Figure 6.
Figure 7.
Knockdown of iASPP expression can strengthen chemosensitivity of HNSCC Tu686 cell line to paclitaxel. CON, shRNA-NC and shRNA-iASPP cells were treated with increasing concentrations of paclitaxel for 48 h, and the cell survival rate was assessed by CCK-8 assays. Results represent the mean ± SD of three independent experiments."
Figure 7.
Figure 8.
The effect of knockdown of iASPP expression on cell apoptosis of Tu686 cells treated with paciltaxel for 24 hours was analyzed by flow cytometry. Representative histograms showing apoptosis of CON (A), shRNA-NC (B) and shRNA-iASPP cells (C). The apoptosis ratio of shRNA-iASPP cells was significantly higher than that of CON cells and shRNA-NC cells (D) (*P=0.000). Results of graphic D represent the mean ± SD of three independent experiments."
Figure 8.
Figure 9.
The effect of knockdown of iASPP expression on cell cycle distribution of Tu686 cells treated with paciltaxel for 24 hours was analyzed by flow cytometry. Representative histograms showing cell cycle distribution of CON (A), shRNA-NC (B) and shRNA-iASPP cells (C). The percentage of shRNA-iASPP cells in G2/M phase was strikingly increased (*P=0.000), and the percentage of cells in S phase decreased significantly (*P=0.000). Results of graphic D represent the mean ± SD of three independent experiments."
Figure 9.
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