Yue He1, Hui Zhao1, Xing-Ming Li2, Cheng-Hong Yin1, Yu-Mei Wu3. 1. Department of Gynecological Oncology, Beijing Obstetrics and Gynecology Hospital, Capital Medical University, Dongcheng District, Beijing 100006, China. 2. School of Health Management and Education, Capital Medical University, Fengtai District, Beijing 100069, China. 3. Department of Gynecological Oncology, Beijing Obstetrics and Gynecology Hospital, Capital Medical University, Dongcheng District, Beijing 100006, China. Electronic address: wym597118@163.com.
Abstract
AIM: To explore the feasibility of using mesothelin (MESO) as a tumor-associated antigen (TAA) in cervical squamous cell carcinoma and its function in the development of cervical cancer. METHODS: We collected eight cervical tissue samples of squamous cell carcinoma as the test group and eight samples of cervicitis as the control group from patients who underwent a hysterectomy because of a diagnosis of myoma. Then we used western blotting to screen for a potential TAA in cervical squamous cell carcinoma samples. In addition, Lentivirus-mediated RNAi was used to downregulate the expression of the MESO gene (MSLN) in SiHa cells. Fluorescence-activated cell sorting (FACS), 3‑(4,5)-dimethylthiahiazo (-z‑y1)‑3,5‑di‑phenytetrazoliumromide (MTT), and wound healing were used to examine cell apoptosis, cell proliferation, and cell migration respectively. RESULTS: Results of the western blotting showed that the MESO protein expressed highly in the cervical squamous cell carcinoma and paracancerous tissues in contrast to the cervicitis tissue (p = 0.242). We used quantitative PCR to verify that the expression of MSLN was 21.6% in the cells undergoing knockdown compared to that in the control cells, and thus, decided to continue with further experiments. We confirmed by FACS that the apoptosis rate in the SiHa cell group undergoing MSLN knockdown (KD group) was significantly higher than that in negative control (NC) group of SiHa cells (p = 0.014). The proliferation of cells was examined by MTT and the difference between the KD and NC groups was found to be statistically significant (p = 0.002). However, since the difference was <20% we did not consider it to be clinically significant. Cell migration ability was verified by wound healing test and found to be 43% in the KD group and 38% in the NC group after 48 h, but the difference was not statistically significant (p > 0.05). CONCLUSIONS: MESO might be used as a TAA for diagnosing cervical squamous cell carcinoma. When MSLN was knocked down in SiHa cells, cell apoptosis increased, but no significant effects were observed on cell proliferation and migration. Thus, our study shows that MSLN plays a role in the apoptosis of cervical squamous cell carcinoma cells, and since this might affect tumor progression, further research is warranted to understand how MSLN plays this role.
AIM: To explore the feasibility of using mesothelin (MESO) as a tumor-associated antigen (TAA) in cervical squamous cell carcinoma and its function in the development of cervical cancer. METHODS: We collected eight cervical tissue samples of squamous cell carcinoma as the test group and eight samples of cervicitis as the control group from patients who underwent a hysterectomy because of a diagnosis of myoma. Then we used western blotting to screen for a potential TAA in cervical squamous cell carcinoma samples. In addition, Lentivirus-mediated RNAi was used to downregulate the expression of the MESO gene (MSLN) in SiHa cells. Fluorescence-activated cell sorting (FACS), 3‑(4,5)-dimethylthiahiazo (-z‑y1)‑3,5‑di‑phenytetrazoliumromide (MTT), and wound healing were used to examine cell apoptosis, cell proliferation, and cell migration respectively. RESULTS: Results of the western blotting showed that the MESO protein expressed highly in the cervical squamous cell carcinoma and paracancerous tissues in contrast to the cervicitis tissue (p = 0.242). We used quantitative PCR to verify that the expression of MSLN was 21.6% in the cells undergoing knockdown compared to that in the control cells, and thus, decided to continue with further experiments. We confirmed by FACS that the apoptosis rate in the SiHa cell group undergoing MSLN knockdown (KD group) was significantly higher than that in negative control (NC) group of SiHa cells (p = 0.014). The proliferation of cells was examined by MTT and the difference between the KD and NC groups was found to be statistically significant (p = 0.002). However, since the difference was <20% we did not consider it to be clinically significant. Cell migration ability was verified by wound healing test and found to be 43% in the KD group and 38% in the NC group after 48 h, but the difference was not statistically significant (p > 0.05). CONCLUSIONS: MESO might be used as a TAA for diagnosing cervical squamous cell carcinoma. When MSLN was knocked down in SiHa cells, cell apoptosis increased, but no significant effects were observed on cell proliferation and migration. Thus, our study shows that MSLN plays a role in the apoptosis of cervical squamous cell carcinoma cells, and since this might affect tumor progression, further research is warranted to understand how MSLN plays this role.