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Objective:To investigate the biological characteristics of proliferation, apoptosis, migration and invasion of radiation-induced polyploid cervical cancer HeLa cells, and to analyze the potential facilitation of polyploid HeLa cells in cervical cancer recurrence after radiotherapy.Methods:HeLa cells were irradiated by 6 MV-X ray with 7 Gy and 14 Gy, the cells were cultured until the third day, and then they were recorded as 7 Gy group and 14 Gy group respectively. The unirradiated HeLa cells were recognized as the control group. The cell morphology was checked under optical microscope. Flow cytometry was used to determine cell ploidy. MTT assay was applied to detect cell proliferation. Flow cytometry by AnnexinⅤ-FITC/PI double labeling was used to detect apoptosis. The ability of migration and invasion was detected by Transwell assay. The expression levels of STAT3 signal pathway-related proteins were analyzed by Western blotting.Results:Compared with the control group, the volume of HeLa cells in 7 Gy group and 14 Gy group increased significantly. The percentages of polyploid HeLa cell subsets in the control group, 7 Gy group and 14 Gy group were (6.33±1.26) %, (21.13±0.50) % and (46.07±1.68) % respectively, with a statistically significant difference ( F=780.47, P<0.001) . The absorbance values in the control group, 7 Gy group and 14 Gy group of polyploidy HeLa cells were 0.21±0.01, 0.23±0.02, 0.16±0.01 at 24 h, 0.37±0.03, 0.38±0.06, 0.21±0.00 at 48 h, 0.66±0.02, 0.55±0.01, 0.28±0.01 at 72 h, and there were statistically significant differences ( F=31.62, P=0.001; F=20.10, P=0.002; F=708.52, P<0.001) . Further pairwise comparison showed that the proliferation abilities of polyploidy HeLa cells of the 14 Gy group at 24, 48 and 72 h were significantly lower than those of the control group and the 7 Gy group (all P<0.05) . The proportions of apoptotic cell subset in the control group, 7 Gy group and 14 Gy group were (3.67±1.16) %, (3.07±0.81) %, (3.83±0.91) %, the proportions of early apoptotic subset were (2.33±0.35) %, (2.13±0.61) %, (2.23±0.32) %, and the proportions of late apoptotic subset were (1.33±0.81) %, (0.93±0.31) %, (1.60±0.60) % respectively. There were no statistically significant differences ( F=0.52, P=0.620; F=0.15, P=0.864; F=0.92, P=0.450) . The migrated numbers of cells in the control group, 7 Gy group and 14 Gy group were 297.40±26.53, 121.33±15.16, 18.40±4.79, and the invaded numbers were 195.67±20.26, 63.60±6.91, 9.47±3.23 respectively, with statistically significant differences ( F=647.28, P<0.001; F=213.94, P<0.001) . Compared with the control group, the migration and invasion abilities of polyploid HeLa cells in the 7 Gy and the 14 Gy groups were significantly decreased, and the migration and invasion abilities of polyploid HeLa cells in the 14 Gy group were significantly lower than those in the 7 Gy group (all P<0.001) . The expression levels of P-STAT3 (Tyr 705) and Bcl-2 in radiation-induced polyploidy HeLa cells were higher than those in the control group, and the expression levels were further increased with the increase of radiation dose. Compared with the control group, the expression levels of Survivin and Mcl-1 in polyploid HeLa cells in the 14 Gy group were up-regulated (both P<0.05) . There was no significant difference in Bcl-xL expression among the three groups ( F=0.52, P=0.618) . Conclusion:The proliferation, migration and invasion abilities of polyploid HeLa cells are reduced by radiation, and the proportion of apoptotic subset is not significantly changed, but the activation of STAT3 signaling pathway is accompanied by up-regulation of downstream anti-apoptotic related proteins, which is favorable for the polyploid tumor cells to be the potential risk factor of recurrent cervical cancer after radiotherapy.
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Background The incidence of dry eye is increasing among young adults because of wide usage of video display terminal.But the early diagnosis of dry eye still presents challenge to medical practitioners.The accurate diagnosis and treatment of the dry eye,therefore,is a topic of high interest to researchers.Previous examination outcome of dry eye is interferred primarily due to invasive procedure.It is very important to search an examination approach.Objective This study was to use Keratograph 5M,a non-invasive ocular surface analyzer to evaluate the influence of watching video display terminal on ocular surface and tear film.Methods Eighty-one eyes of 81 health volunteers among 18-30 years were enrolled in Affiliated Eye Hospital of Nanchang University from March 1,2015 to November 10,2015 under the informed consent,including 39 males and 42 females.The subjects watched the computer for continuously 3 hours under the nature light,and ocular surface related examinations were performed and compared before and after video display terminal exposure,including non-invasive tear film break-up time (NITBUT),tear meniscus height,conjunctival hyperemia scoring,limbal congestion scoring,corneal fluorescein staining scoring,meibomian gland imaging and lipid layer analysis.Results The number of eyes with visual fatigue,dryness,pain,blurring and conjunctival congestion was significantly increased after 3-hour video display terminal exposure in comparison with before (all at P＜0.01).The initial NITBUT and mean NITBUT were (6.086± 3.701) s and (9.103 ± 4.680) s,and tear meniscus height was (0.190 ± 0.032) mm after trail,which were significantly lower than (11.445 ±4.964) s,(14.626 ±4.467) s and (0.212 ±0.040) mm of before trail,respectively;The conjunctical hyperemia scoring and limbal congestion scoring were 0.869 ±0.311 and 0.572 ±0.276 after trial,which were significantly higher than 0.780 ± 0.306 and 0.509 ± 0.266 before trail,showing significant differences before and after exposure of video display terminal (all at P＜0.01).The intraocular pressure and the eye number of different scores of corneal fluorescence staining,abnormal meibomian gland and different morphological lipid layer of tear were unchanged before and after exposure of video display terminal.Conclusions Long-term exposure of video display terminal results in significant and temporary adverse influence on tear film and ocular surface.Keratograph 5M non-invasive ocular sudace analyzer can objectively assess overall ocular surface conditions.