| Part One: The investigate of NALP3 inflammsome and the inflammsome related factor in esophageal squamous cell carcinoma.Objective: To investigate the expression of NACHT-LRR-PYD containing proteins 3(NALP3), apoptosis associated speck-like protein containing a card(ASC), Caspase-1 and the inflammasome related factor IL-1β and TNF-α in esophageal squamous cell carcinoma(ESCC).Methods: The expression of IL-1β and TNF-α were tested by Enzyme linked immunosorbent assay(Elisa), m RNA of NALP3, ASC and caspase-1 were detected by real-time quantitative polymerase chain reaction(RT-q PCR), the protein of NALP3, ASC and caspase-1 in esophageal squamous cell carcinoma were examined with Western-Blot.Result: The secreted of IL-1β and TNF-α in esophageal squamous cancer were higher than those in normal esophageal tissue. [IL-1β,(3.84±0.78) pg/ml vs.(0.27±0.04) pg/ml,(P<0.01); TNF-α,(1.07±0.13) pg/ml vs.(0.40±0.06) pg/ml,(P<0.05)]. At the same time, The expression of NALP3 m RNA, Caspase-1 m RNA, ASC m RNA were higher in tumor tissue. [NALP3 m RNA,(1.06±0.14 vs. 0.89±0.12),(P<0.01); Caspase-1 m RNA,(0.93±0.09 vs. 0.80±0.12),(P<0.05); ASC m RNA,(0.98±0.08 vs. 0.85±0.15),(P<0.05)]. The expression of protein NALP3, Caspase-1, ASC were higher in tumour tissue. [NALP3,(1.48±0.04 vs. 1.09±0.08),(P<0.05); Caspase-1,(0.59±0.03 vs. 0.36±0.04),(P<0.01); ASC,(0.42±0.10 vs. 0.18±0.08),(P<0.05)].Conclusion: The expression of NALP3 inflammsome and inflammasome related factor IL-1β and TNF-α were higher in tumour tissue than normal tissue. Part Two: Effects of IL-1β on proliferation and migration of ESC410 cell lineObjective: To study the proliferation and migration ability of human esophageal squamous cell line of high incidence area of esophageal cancer Si Chuan(ESC410) treated with different leve of IL-1β.Methods: The proliferation of ESC410 was evaluated by EDU(5-Ethynyl- 2’- deoxyuridine) essay. Cells migration was examined by scratch test.Results: Treated with 1ng/ml IL-1β, the ESC410 proliferation radio was higher than control group [24h,(42.6±1.0)% vs.(37.8±2.3)%,(P<0.05)], and migration distance(fx) was higher than control group also. [(12h,(219.1±13.8) vs.(121.4±12.0),(P<0.01); 24 h,(416.5±21.3) vs.(192.7±16.8),(P<0.01)]. Treated with 10ng/ml IL-1β, the ESC410 proliferation radio was no difference with the control group [24h,(38.9±1.4)% vs.(37.8±2.3)%,(P>0.05)], and migration distance(fx) was no difference with the control group also. [12h,(135.5±35.2) vs.(121.4±12.0),(P>0.05); 24 h,(200.9±18.5) vs.(192.7±16.8),(P>0.05)].Conclusion: IL-1β had positive effects on ESC410, not only increasing proliferation but also enhancing migration abillty, at leve of 1ng/ml, but at leve of 10ng/ml, the effects above was disappear. |
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