ROLE OF INTEGRIN ALPHA 6 SINGNALING PATHWAY IN NCI-H460 NON-SMALL CELL LUNG CARCINOMA GROWTH AND METASTASIS
(0) Objectives Complete resection of early-stage nonsamall cell lung cancer (NSCLC) is potentially curative, yet approximately 50％ of patients are at risk for developing metastatic recurrence. We previously suppressed integrin a6(J4 expression in non-small cell lung cancer cells by RNA interference. In the percent study we investigated the role of integrin a6 signaling in the growth and metastasis of non-small cell lung cancer in vitro and in vivo xenograft nude-rat model. Methods we stably silenced the integrina6P4 expression in highly metastic NCIH460SM cells via lentivirus-mediated transduction. We verified the expression of a6β4 integrin by qRT-PCR and western blot, and observed the cellular morphology by microscope. Tumor cells growth was detected by cell counting, MTT and MTS assay in vitro. Furthermore, the changes of cell cycle and apoptosis were analyzed by flow cytometry. Anchorageindependent colony formation was detected by soft agar assay, and cell invasive potential was determined by invasion assay, seven cell lines including NCI-H460, its variant cell line H460SM , H460SM-68 or H460SM-71 which had been knocking out P4 gene, H460SM-75 or H460SM-76 which had been silenced a6 gene were cultured at 37 °C 5％CO2 in RPMI1640 medium containing 10％ fetal bovine serum(FBS). 56 nude rats were divided into 7 groups. They were injected different cell lines (NCI-H460, H460SM, H460SM-68, H460SM-71, H460SM-75, and H460SM-76). 1 × 106 Cells were inoculated into the left oxter of nude rat to establish the xenograft model. Weight the nude rats and measurethe tumor every other day to compare the tumor growth of every group. Anatomize nude rats and observe the metastasis on heart, liver, spleen, lung, kidney and lymph node. Tissues and organs were fixed in 10％ buffered formalin for histologic examination. Results we silenced successfully the integrina6β4 expression in highly metastic NCI-H460SM cells via lentivirus-mediated transduction. shfW H460SM transduced stable cells showed decreased growth rate compared with that of control nontargeting shRNA cells. The sha6f)4 H460SM cells showed a decrease in anchorage-independent colony formation, mobility and invasiveness. In contrast, there was no statistically significant difference in apoptotic rates among all of those non-small cell lung cancer cells (H460SM-NS, H460SM-68, H460SM -71, H460SM-75, H460SM-76). The distribution ofsh04 H460SM transduced stable cells in the G0/G1 phase was significantly higher than that of control nontargeting shRNA suggesting that the cell proliferation was inhibited by blocking the cell cycle in G0/G1 phase (P=0.017, P=0.013 respectively). PI value of shp4-H460SM transduced stable cells was lower than that of H460SM-NS, suggesting the proliferation of shRNA-fW transfected cells was inhibited. Consistent with our in vitro findings, sha6|34 H460SM cells demonstrated enhanced tumorgenity ( P=0.025 ) and metastasis (P=0.040) in xerograft nude mice. Conclusions Our data indicated that integrin a6 signaling may play an important role in NSCLC growth and metastasis. Understanding the mechanism of this signaling in NSCLC growth and metastasis may develop novel strategies for early stage NSCLC diagnosis and treatment.
Xenograft model NSCLC Integrin a6β4 Growth Metastasis
Chen Yan Wang Xi Cai Wu Zhi Ping Jin Cong Guo Liu Xin Zhou Yong Chun Chen Xiao Qun Li Jia Ming-Sound Tsao
Yunnan Tumor Institute, The Third Affiliated Hospital of Kunming Medical University (Tumor Hospital YunnanTumor Institute, The Third Affiliated Hospital of Kunming Medical University (Tumor Hospital o Ontario Cancer Institute and Princess Margaret Hospital, University Health Network, Toronto,Ontario,
国际会议
亚太地区国际肿瘤生物学和医学学术会议、全国肿瘤标志学术大会、第六届中国中青年肿瘤专家论坛
上海
英文
404-405
2011-10-13（万方平台首次上网日期，不代表论文的发表时间）