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Objective To investigate the expression levels of serum exosomes integrin α5, β5 and β6 and their clinical significance in patients with different stages of colorectal cancer (CRC). Methods The serum samples were collected from 50 patients with CRC admitted in the Affiliated Hospital of Hunan Academy of Traditional Chinese Medicine and Hunan Cancer Hospital from January 2016 to October 2020, and were divided into CRC stage Ⅰ group (n=5), CRC stage Ⅱ group (n=19), CRC stage Ⅲ group (n=14), and CRC stage Ⅳ group (only liver metastasis, n=12) according the TNM stage. The serum samples were collected from 10 healthy people who underwent physical examination in the Health Management Center of the Affiliated Hospital of Hunan Academy of Traditional Chinese Medicine from August 2018 to May 2020, and set as healthy people group. The exosomes were purified, and then identified by nanoparticle tracking assay (NTA) and Western blotting. The expression levels of integrin α5,β5 and β6 were detected by Western blotting, the survival time of 50 patients with CRC were followed up. Kaplan-Meier method was used to draw the survival curve, and Cox proportional hazards model was used for survival analysis. Results The 50 patients with CRC [26 males (52.0%) and 24 females (48.0%)] are mainly middle- and elderly-aged with average age of 58 years. Five cases (10.0%)in TNM stage Ⅰ, 19 cases (38.0%) in stage Ⅱ, 14 cases (28.0%) in stage Ⅲ and 12 cases (24.0%) in stage Ⅳ (only liver metastasis).Up to February 2021, the follow-up time was 5-60 months, and the median follow-up time was 44 months, no cases fell off or lost follow-up. NTA analysis showed that the particle diameter of serum exosomes in all serum samples were concentrated in 50-150 nm.Western blotting results showed that the marker proteins of serum exosomes ALIX and HSP70 were expressed in all serum samples,which confirmed that these extracts were exosomes. Integrin α5, β5 and β6 were expressed in different degrees, the expression levels of integrin α5 and β5 increased with the increase of TNM stage, and significantly higher in live metastasis than in non-stage Ⅳ(P<0.05). However, no significant relationship existed between the expression of integrin β6 and TNM stage (P>0.05). Kaplan-Meier method showed that integrin α5, β5 and TNM stage were related with survival of patients with CRC (P<0.05). The results of Cox proportional risk model suggested that integrin α5 and the degree of pathological differentiation were independent influencing factors of survival in patients with CRC (P<0.05). Conclusion Exosomes integrin α5, β5 are related to liver metastasis and TNM stage of CRC. Exosomes integrin α5 and the degree of pathological differentiation are related to the prognosis of colorectal cancer,and integrin α5 and β5 may become potential serum markers.
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| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
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