Preliminary Study of the Role of INPP4B in Promoting Colorectal Cancer Metastasis and the Mechanisms Involved

LAI Meng, MAO Zhigang, TANG Deng, LAN Siqi, YAN Ruiting, XIANG Qi, ZHAO Xianxian, SU Mi, WANG Yufang

Abstract

To investigate the expression of inositol polyphosphate 4-phosphatase type Ⅱ B (INPP4B) in colorectal cancer (CRC) and the relevant clinical significance, to determine the relationship between INPP4B and matrix metallopeptidase 7 (MMP7) in CRC cells, and to make preliminary exploration of the effects of INPP4B on the proliferation and migration of CRC cells and mechanisms involved.

Methods 

 The TIMER2.0 and GEPIA2 databases were used to analyze the differences in INPP4B expression between cancer and para-cancerous tissues and the effects of such differences on the prognosis of CRC. The expression of INPP4B in 102 surgically resected CRC tumors was determined by immunohistochemistry (IHC), and the correlation between INPP4B and clinical pathological indicators was analyzed. In CRC cells with overexpressed/knocked-down INPP4B, the expression of INPP4B and MMP7 were examined by real time fluorogenic quantitative PCR, the protein expression of INPP4B was assessed by Western blot, cell proliferation was determined using the CellTiter 96® AQueous One assay, and cell migration and invasion were assessed using wound healing assay and real-time label-free dynamic cell analysis (RTCA). The LinkedOmics database was used to analyze signaling pathways related to INPP4B function, and the role of potential key molecules was validated at the cellular level.

Results 

Analysis with the TIMER2.0 database and GEPIA2 database showed elevated INPP4B expression (colon adenocarcinoma [COAD]: 2.30, rectal adenocarcinoma [READ]: 2.33) in CRC compared to normal tissue (COAD: 1.91, READ: 1.89). IHC testing confirmed that INPP4B was upregulated in clinical CRC tissues and paracancerous tissues (P<0.001). Cox regression model analysis showed that INPP4B (hazards ratio [HR]=1.457, 95% confidence interval [CI]: 1.003-2.115) affected the prognosis of CRC, and the Kaplan-Meier curve showed that patients with high INPP4B expression had shorter overall survival (P<0.05). χ2 test was performed to analyze the relationship between INPP4B expression and clinicopathological indexes, and it was found that high expression of INPP4B was correlated with lymph node metastasis (χ2=3.997, P=0.046) and neural invasion(χ2=8.511, P=0.004). In in vitro experiments, CRC cells overexpressing INPP4B showed a significantly increased cell proliferation and migration compared to the cells in the control group (P<0.05). Analysis using the LinkedOmics database showed that INPP4B was correlated with extracellular matrix remodeling and cell migration. Pearson's correlation analysis showed that MMP7 was positively correlated with INPP4B (r=0.3782, P<0.001). INPP4B overexpression or knockdown in vitro also led to the upregulation or the downregulation of MMP7 expression in CRC cells.

Conclusion 

 INPP4B is highly expressed in CRC tissues and significantly correlated with lymph node metastasis, neural invasion, and patient prognosis. MMP7 may mediate the role of INPP4B in promoting CRC cell migration and invasion.

 

 Keywords: INPP4B, Colorectal cancer, MMP7

 

Full Text:

PDF


References


SUNG H, FERLAY J, SIEGEL R L, et al. Global cancer statistics 2020: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J Clin, 2021, 71(3): 209–249. doi: 10. 3322/caac.21660.

ZHENG R, ZHANG S, ZENG H, et al. Cancer incidence and mortality in China, 2016. J Natl Cancer Cent, 2022, 2(1): 1–9. doi: 10.1016/j.jncc.2022. 02.002.

LI N, LU B, LUO C, et al. Incidence, mortality, survival, risk factor and screening of colorectal cancer: a comparison among China, Europe, and northern America. Cancer Lett, 2021, 522: 255–268. doi: 10.1016/j.canlet. 2021.09.034.

BILLER L H, SCHRAG D. Diagnosis and treatment of metastatic colorectal cancer: a review. JAMA, 2021, 325(7): 669–685. doi: 10.1001/jama.2021.0106.

HAMILA S A, OOMS L M, RODGERS S J, et al. The INPP4B paradox: like PTEN, but different. Adv Biol Regul, 2021, 82: 100817. doi: 10.1016/j. jbior.2021.100817.

GUO S T, CHI M N, YANG R H, et al. INPP4B is an oncogenic regulator in human colon cancer. Oncogene, 2016, 35(23): 3049–3061. doi: 10.1038/onc.2015.361.

SUNG J Y, NA K, KIM H S. Down-regulation of inositol polyphosphate 4-phosphatase type Ⅱ expression in colorectal carcinoma. Anticancer Res, 2017, 37(10): 5525–5531. doi: 10.21873/anticanres.11984.

HE L, KANG Q, CHAN K I, et al. The immunomodulatory role of matrix metalloproteinases in colitis-associated cancer. Front Immunol, 2022, 13: 1093990. doi: 10.3389/fimmu.2022.1093990.

SHATTUCK-BRANDT R L, LAMPS L W, HEPPNER GOSS K J, et al. Differential expression of matrilysin and cyclooxygenase-2 in intestinal and colorectal neoplasms. Mol Carcinog, 1999, 24(3): 177–187.

MORI M, BARNARD G F, MIMORI K, et al. Overexpression of matrix metalloproteinase-7 mRNA in human colon carcinomas. Cancer, 1995, 75(6 Suppl): 1516–1519. doi: 10.1002/1097-0142

LI T, FAN J, WANG B, et al. TIMER: a web server for comprehensive analysis of tumor-infiltrating immune cells. Cancer Res, 2017, 77(21): e108–e110. doi: 10.1158/0008-5472.CAN-17-0307.

LI T, FU J, ZENG Z, et al. TIMER2.0 for analysis of tumor-infiltrating immune cells. Nucleic Acids Res, 2020, 48(W1): W509–W514. doi: 10. 1093/nar/gkaa407.

LI B, SEVERSON E, PIGNON J C, et al. Comprehensive analyses of tumor immunity: implications for cancer immunotherapy. Gen Biol, 2016, 17(1): 174. doi: 10.1186/s13059-016-1028-7.

TANG Z, KANG B, LI C, et al. GEPIA2: an enhanced web server for large-scale expression profiling and interactive analysis. Nucleic Acids Res, 2019, 47(W1): W556–W560. doi: 10.1093/nar/gkz430.

VASAIKAR S V, STRAUB P, WANG J, et al. LinkedOmics: analyzing multi-omics data within and across 32 cancer types. Nucleic Acids Res, 2018, 46(D1): D956–D963. doi: 10.1093/nar/gkx1090.

FRUMAN D A, ROMMEL C. PI3K and cancer: lessons, challenges and opportunities. Nat Rev Drug Disc, 2014, 13(2): 140–156. doi: 10.1038/nrd4204.

BRANA I, PHAM N A, KIM L, et al. Novel combinations of PI3K-mTOR inhibitors with dacomitinib or chemotherapy in PTEN-deficient patient-derived tumor xenografts. Oncotarget, 2017, 8: 84659-84670 . doi: 10. 18632/oncotarget.19109.

TSUBAKI M, TAKEDA T, NOGUCHI M, et al. Overactivation of Akt contributes to MEK inhibitor primary and acquired resistance in colorectal cancer cells. Cancers (Basel), 2019, 11(12): 1866. doi:10.3390/cancers11121866.

LIU H, PADDOCK M N, WANG H, et al. The INPP4B tumor suppressor modulates EGFR trafficking and promotes triple-negative breast cancer. Cancer Discov, 2020, 10(8): 1226–1239. doi: 10.1158/2159-8290.CD-19-1262.

FEDELE C G, OOMS L M, HO M, et al. Inositol polyphosphate 4-phosphatase Ⅱ regulates PI3K/Akt signaling and is lost in human basal-like breast cancers. Proc Natl Acad Sci U S A, 2010, 107(51): 22231–22236. doi: 10.1073/pnas.1015245107.

Li CHEW C, LUNARDI A, GULLUNI F, et al. In Vivo role of INPP4B in tumor and metastasis suppression through regulation of PI3K-AKT signaling at endosomes. Cancer Discov, 2015, 5(7): 740–751. doi: 10.1158/ 2159-8290.CD-14-1347.

HODGSON M C, SHAO L J, FROLOV A, et al. Decreased expression and androgen regulation of the tumor suppressor gene INPP4B in prostate cancer. Cancer Res, 2011, 71(2): 572–582. doi: 10.1158/0008-5472.CAN-10-2314.

HODGSON M C, DERYUGINA E I, SUAREZ E, et al. INPP4B suppresses prostate cancer cell invasion. Cell Commun Sign, 2014, 12: 61. doi: 10.1186/s12964-014-0061-y.

CHAURASIYA S, WU W, STROM A M, et al. Estrogen receptor β regulates AKT activity through up-regulation of INPP4B and inhibits migration of prostate cancer cell line PC-3. Proc Natl Acad Sci U S A, 2020, 117(42): 26347–26355. doi: 10.1073/pnas.2007160117.

ZHANG F, LI J, ZHU J, et al. IRF2-INPP4B-mediated autophagy suppresses apoptosis in acute myeloid leukemia cells. Biol Res, 2019, 52(1): 11. doi: 10.1186/s40659-019-0218-7.

DZNELADZE I, HE R, WOOLLEY J F, et al. INPP4B overexpression is associated with poor clinical outcome and therapy resistance in acute myeloid leukemia. Leukemia, 2015, 29(7): 1485–1495. doi: 10.1038/leu. 2015.51.


Refbacks

  • There are currently no refbacks.