Different Proteins Regulated Apoptosis, Proliferation and Metastasis of Lung Adenocarcinoma After Radiotherapy at Different Time
Different Proteins Regulated Apoptosis, Proliferation and Metastasis of Lung Adenocarcinoma After Radiotherapy at Different Time
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不同时间点不同蛋白调控肺腺癌放疗后的细胞凋亡、增殖和转移
DOI：
10.2147/cmar.s219967
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发表时间：
2020-01-01
影响因子：
3.3
通讯作者：
Li, W. H.
中科院分区：
文献类型：
--
作者：
Dai, P. L.;Du, X. S.;Li, W. H.
引言肺癌细胞辐射后的生物学变化对于减少肺癌的复发和转移具有重要意义。为了优化肺腺癌的放射治疗，我们的研究系统地探讨了放射后残留的A549和XWLC-05细胞的生物学行为机制。方法采用集落形成实验、细胞增殖实验、细胞迁移实验、流式细胞术、BALB/C-nu小鼠异种移植模型以及pan-AKT、p-Akt380、p-Akt473、PCNA、DNA-PKCS、KU70、KU80、CD133、CD144、MMP2和P53的蛋白质印迹来评估0、4和8 Gy辐照后的生物学变化。体外照射后0-336小时，移植组20Gy，照射移植组，体内残留肿瘤0、7、14、21和28天组。结果体内外放射后残留的XWLC-05细胞的细胞增殖能力和放射敏感性均优于A549细胞。 MMP-2在体外和体内具有统计学差异，并且随着细胞体外迁移能力的增加而增加。 XWLC-05和A549细胞中PCNA和P53有统计学差异，其变化与体外照射后336小时内残留细胞的增殖相似。体内Pan-AKT在照射后增加，残留肿瘤21天组（1.5722）与移植组（0.9763，p=0.018）和照射移植组（0.8455，p=0.006）之间有统计学差异。当残留肿瘤达到0.5 mm2后21天，Pan-AKT升至最高。 MMP2在移植组（0.4619）和残留肿瘤14天组（0.8729，p=0.043）之间具有统计学差异。 P53在残瘤7天组（0.6184）和残瘤28天组（1.0394，p=0.007）之间有统计学差异。 DNA-PKCS在残瘤28天组（1.1769）与移植组（0.2483，p=0.010）、照射移植组（0.1983，p=0.002）与残瘤21天组（0.2017，p=0.003）、残瘤0天组（0.5992）与照射移植组之间具有统计学差异(0.1983，p=0.027)和残留肿瘤21天组(0.2017，p=0.002)。 KU80和KU70在任何时间点都没有统计差异。结论不同蛋白在不同时期对肺腺癌放疗后的细胞凋亡、增殖和转移具有调节作用。 MMP-2可能调节XWLC-05和A549细胞的体外和体内转移能力。 PCNA 和 P53 可能在体外辐射后 336 小时内的体外 XWLC-05 和 A549 细胞增殖中发挥重要作用。此后，P53可能通过PI3K/AKT途径调节体外照射后的细胞增殖。体外336小时后，DNA-PKCS可能比KU70和KU80在DNA损伤修复中发挥更重要的作用，因为它在辐照后比KU70和KU80迅速上升。不同细胞在放疗后凋亡、增殖和转移的时间节律不同。应注意照射后细胞的时间节律，并注意抵抗癌细胞的增殖和转移。
Introduction The biological changes after irradiation in lung cancer cells are important to reduce recurrence and metastasis of lung cancer. To optimize radiotherapy of lung adenocarcinoma, our study systematically explored the mechanisms of biological behaviors in residual A549 and XWLC-05 cells after irradiation. Methods Colony formation assay, cell proliferation assay, cell migration assay, flow cytometry, BALB/C-nu mice xenograft models and Western blot of pan-AKT, p-Akt380, p-Akt473, PCNA, DNA-PKCS, KU70, KU80, CD133, CD144, MMP2 and P53 were used in our study to assess biological changes after irradiation with 0, 4 and 8 Gy at 0–336 hr after irradiation in vitro and 20 Gy at transplantation group, irradiated transplantation group, residual tumor 0, 7, 14, 21, and 28 days groups in vivo. Results The ability of cell proliferation and radiosensitivity of residual XWLC-05 cells was better than A549 cells after radiation in vivo and in vitro. MMP-2 has statistical differences in vitro and in vivo and increased with the migratory ability of cells in vitro. PCNA and P53 have statistical differences in XWLC-05 and A549 cells and the changes of them are similar to the proliferation of residual cells within first 336 hr after irradiation in vitro. Pan-AKT increased after irradiation, and residual tumor 21-day group (1.5722) has statistic differences between transplantation group (0.9763, p=0.018) and irradiated transplantation group (0.8455, p=0.006) in vivo. Pan-AKT rose to highest when 21-day after residual tumor reach to 0.5 mm2. MMP2 has statistical differences between transplantation group (0.4619) and residual tumor 14-day group (0.8729, p=0.043). P53 has statistical differences between residual tumor 7-day group (0.6184) and residual tumor 28 days group (1.0394, p=0.007). DNA-PKCS has statistical differences between residual tumor 28 days group (1.1769) and transplantation group (0.2483, p=0.010), irradiated transplantation group (0.1983, p=0.002) and residual tumor 21 days group (0.2017, p=0.003), residual tumor 0 days group (0.5992) and irradiated transplantation group (0.1983, p=0.027) and residual tumor 21 days group (0.2017, p=0.002). KU80 and KU70 have no statistical differences at any time point. Conclusion Different proteins regulated apoptosis, proliferation and metastasis of lung adenocarcinoma after radiotherapy at different times. MMP-2 might regulate metastasis ability of XWLC-05 and A549 cells in vitro and in vivo. PCNA and P53 may play important roles in proliferation of vitro XWLC-05 and A549 cells within first 336 hr after irradiation in vitro. After that, P53 may through PI3K/AKT pathway regulate cell proliferation after irradiation in vitro. DNA-PKCS may play a more important role in DNA damage repair than KU70 and KU80 after 336 hr in vitro because it rapidly rose than KU70 and KU80 after irradiation. Different cells have different time rhythm in apoptosis, proliferation and metastasis after radiotherapy. Time rhythm of cells after irradiation should be delivered and more attention should be paid to resist cancer cell proliferation and metastasis.