| PRR11(Proline-rich protein11) is a novel recently identified geneinvolved in both cell cycle regulation and tumorigenesis. Ska2(spindle andkinetochore associated complex subunit2), also known as FAM33A(family with sequence similarity33, member A), is also a recentlyidentified gene involved in both cell cycle regulation and tumorigenesis, itsfunction in lung cancer remains unclear. Our previously study indicatedthat both of them are closely located in17q23, share a bidirectionalpromoter with a unique head-to-head way, and the gene pairs function as aentire and unique transcription unit regulated by NFYB and p53. Thepresent study were designed to investigate the expression and function ofSka2, PRR11-Ska2gene pairs in lung cancer.(1) Expression and clinical significance of Ska2in lung cancerReal time-PCR was used to analyze the expression of Ska2in normaland cancerous lung tissue. The results showed that its expression isupregulated in lung cancer tissue as compared with normal lung tissue, andis positively correlated with the grades and the clinical pathological stage.We also analyzed the prognostic value of Ska2in lung cancer bybioinformatic approaches. The results showed that high Ska2expression is significantly correlated with poor prognosis in lung cancer patients.(2) Expression and clinical significance of PRR11-Ska2“gene pairs”in lung cancerWe analysed the expression correlation of Ska2with PRR11invarious human normal tissues, lung cancer tissues and lung cancermicroarray dataset GSE13213. The results indicated that PRR11expressionis correlated with Ska2expression in all the aforementioned data. Theexpression levels of PRR11and Ska2are also correlated positively withNFYB, and their expression levels are higher in wild type p53lung cancergroup as compared with the mutant p53group. The results stronglysuggested that Ska2and PRR11function as an entire transcriptional unitregulated by p53and NFYB.We analyzed the prognostic value of PRR11-Ska2gene pairs bybioinformatic approaches. The results showed that, compared with eitherhigh expression of PRR11or Ska2, high expression of both PRR11andSka2is more significantly correlated with poorer prognosis in lung cancerpatients. These results indicated that, compared with the single PRR11orSka2gene marker, RR11-Ska2gene pairs is a more sensitive prognosticmarker for lung cancer.(3) Functional analysis of PRR11-Ska2“gene pairs” in lung cancerThe stable cell strains with PRR11and/or Ska2depletion wereestablished using lung cancer cell line A549and H1299by lentivirus-mediated RNA interference. These stable cell strains were namedas LV-shRNA-NC, LV-shRNA-PRR11, LV-shRNA-Ska2andLV-shRNA-P+S, respectively. Western blotting results showed thatexpression level of PRR11and/or Ska2were efficiently silenced in thecorresponding stable cell strains.The phenotypic analysis of the stable cells indicated that the ability ofproliferation, migration and invasion of LV-shRNA-Ska2,LV-shRNA-PRR11, LV-shRNA-P+S were significantly lower thanLV-shRNA-NC in both A549and H1299cells. Notably, compared withLV-shRNA-Ska2and LV-shRNA-PRR11, LV-shRNA-P+S cells showed amore remarkable decrease in proliferation, migration and invasion in bothA549and H1299cells. The results suggested that both PRR11and Ska2arenecessary for tumorigenesis and progression of lung cancer, and the twogenes might work in a synergistic or complementary manner.We further determined the expression changes of cell cycle-andtumor-related genes (CCNA1、CDC2、MAP4K4、RRM1、EPB41L3、CCNA2、NFIB、DHRS2) in the aforementioned stable cells. The resultsindicated that the expression of several genes were differentially changedafter PRR11and/or Ska2depletion, suggesting that PRR11and Ska2arefunctionally correlated, their molecular mechanisms might be different.In summary, our present study demonstrated that Ska2is a novel geneinvolved lung tumorigenesis, and PRR11-Ska2gene pairs are necessary for lung cancer development and serve as a novel potential target for thetreatment of lung cancer. |
PDF Full Text Request