Study On The Chemotherapy Of Laryngeal Squamous Carcinoma By Trichsanthin

Objective:To investigated TCS with or without cisplatin inhibited of Laryngeal Carcinoma (HEp-2and AMC-HN-8cells) and the relationship between the inhibition activity of TCS and its activation of JNK/MAPK. Methods:HEp-2and AMC-HN-8cells were treated with different concentration of TCS, and then were divided into low level cisplatin(3μg/ml), high level cisplatin(10μg/ml), TCS(5μg/ml),the combination group(5μg/ml TCS and3μg/ml cisplatin) and control group. After successive five days, different experiment groups were detected by CCK-8and supernatant was detected by LDH kit. Flow cytometric assay were done to detect the HEp-2and AMC-HN-8cell apoptosis and cell cycle. Realtime RT-PCR was performed to detect P27, P21and some other cycle related genes in HEp-2and AMC-HN-8cells. Western blot was performed to detect changes in JNK, phosphor-JNK, p38, phosphor-p38, NF-κB, I-κB, phospho-I-κB.Results:TCS significantly inhibit cell viability of HEp-2and AMC-HN-8cells, independent of necrosis. TCS can induce HEp-2and AMC-HN-8cell apoptosis and increase the percentage of S phase cell cycle.JNK/MAPK was activated by TCS. The cell viability inhibition and apoptosis activity of TCS can be suppressed by JNK/MAPK inhibitor SP600125. Compared with TCS(5μg/ml) or cisplatin(3μg/ml) alone, the combination of TCS and cisplatin had more effect on inhibition of HEp-2and AMC-HN-8cells. So dose apoptosis rate. The combination group did not add cytotoxicity (P>0.05).While the toxicity of the combination group is much lower than10μg/ml cisplatin(P<0.01).Western blot showed the expression of p-JNK/SAPK significantly increased after treated with5μg/ml TCS for48hours. The expression of NF-κB and phospho-I-KB increased after treated with3μg/ml cisplatin. When5μg/ml TCS combined with3μg/ml cisplatin to treat with HEp-2or AMC-HN-8,the expression of p-JNK/MAPK stay high level and NF-κB and phospho-I-κB decreased dramatically compared with3μg/ml cisplatin alone. Conclusions:These results suggest that trichosanthin could induce apoptosis of laryngeal squamous carcinoma cells (HEp-2and AMC-HN-8) and cell cycle arrest through JNK/MAPK. While trichosanthin could enhance cisplatin-induced apoptosis in HEp-2and AMC-HN-8,at least in part, by inhibiting the NF-κB signaling pathway and activating JNK/SAPK signaling pathway and thus strengthening the antitumor effects of cisplatin,which highlights the possibility of combined drug application of TCS and cisplatin in clinical treatment of laryngocarcinoma.