Microgravity has been shown to impose various effects on breast cancer cells. We exposed human breast cancer cells to simulated microgravity and studied morphology and alterations in gene expression.
Human breast cancer cells were exposed to simulated microgravity in a random positioning machine (RPM) for 24 h. Morphology was observed under light microscopy, and gene alteration was studied by qPCR.
After 24 h, formation of three-dimensional structures (spheroids) occurred. BRCA1 expression was significantly increased (1.9×, p < 0.05) in the adherent cells under simulated microgravity compared to the control. Expression of KRAS was significantly decreased (0.6×, p < 0.05) in the adherent cells compared to the control. VCAM1 was significantly upregulated (6.6×, 2.0×, p < 0.05 each) in the adherent cells under simulated microgravity and in the spheroids. VIM expression was significantly downregulated (0.45×, 0.44×, p < 0.05 each) in the adherent cells under simulated microgravity and in the spheroids. There was no significant alteration in the expression of MAPK1, MMP13, PTEN, and TP53.
Simulated microgravity induces spheroid formation in human breast cancer cells within 24 h and alters gene expression toward modified adhesion properties, enhanced cell repair, and phenotype preservation. Further insights into the underlying mechanisms could open up the way toward new therapies.
Aleshcheva G , Sahana J , Ma X , Hauslage J , Hemmersbach R , Egli M , Infanger M , Bauer J , Grimm D : Changes in morphology, gene expression and protein content in chondrocytes cultured on a random positioning machine. PLoS One 8, e79057 (2013)
Aleshcheva G , Wehland M , Sahana J , Bauer J , Corydon TJ , Hemmersbach R , Frett T , Egli M , Infanger M , Grosse J , Grimm D : Moderate alterations of the cytoskeleton in human chondrocytes after short-term microgravity produced by parabolic flight maneuvers could be prevented by up-regulation of BMP-2 and SOX-9. FASEB J. 29, 2303–2314 (2015)
Arizti P , Fang L , Park I , Yin Y , Solomon E , Ouchi T , Aaronson SA , Lee SW : Tumor suppressor p53 is required to modulate BRCA1 expression. Mol. Cell. Biol. 20, 7450–7459 (2000)
Blaber EA , Finkelstein H , Dvorochkin N , Sato KY , Yousuf R , Burns BP , Globus RK , Almeida EA : Microgravity reduces the differentiation and regenerative potential of embryonic stem cells. Stem Cells Dev. 24, 2605–2621 (2015)
Boulton SJ : Cellular functions of the BRCA tumour-suppressor proteins. Biochem. Soc. Trans. 34, 633–645 (2006)
Chen ZY , Guo S , Li BB , Jiang N , Li A , Yan HF , Yang HM , Zhou JL , Li CL , Cui Y : Effect of weightlessness on the 3D structure formation and physiologic function of human cancer cells. Biomed. Res. Int. 2019, 4894083 (2019)
Easton D , Ford D , Peto J : Inherited susceptibility to breast cancer. Cancer Surv. 18, 95–113 (1993)
Feger BJ , Thompson JW , Dubois LG , Kommaddi RP , Foster MW , Mishra R , Shenoy SK , Shibata Y , Kidane YH , Moseley MA , Carnell LS , Bowles DE : Microgravity induces proteomics changes involved in endoplasmic reticulum stress and mitochondrial protection. Sci. Rep. 6, 34091 (2016)
Froehlich K , Haeger JD , Heger J , Pastuschek J , Photini SM , Yan Y , Lupp A , Pfarrer C , Mrowka R , Schleussner E , Markert UR , Schmidt A : Generation of multicellular breast cancer tumor spheroids: comparison of different protocols. J. Mammary Gland Biol. Neoplasia. 21, 89–98 (2016)
Grimm D , Infanger M , Westphal K , Ulbrich C , Pietsch J , Kossmehl P , Vadrucci S , Baatout S , Flick B , Paul M , Bauer J : A delayed type of three-dimensional growth of human endothelial cells under simulated weightlessness. Tissue Eng. Part A 15, 2267–2275 (2009)
Hada M , Ikeda H , Rhone JR , Beitman AJ , Plante I , Souda H , Yoshida Y , Held KD , Fujiwara K , Saganti PB , Takahashi A : Increased chromosome aberrations in cells exposed simultaneously to simulated microgravity and radiation. Int. J. Mol. Sci. 20, 43 (2018)
Hendrix A , Hume AN : Exosome signaling in mammary gland development and cancer. Int. J. Dev. Biol. 55, 879–887 (2011)
Huang B , Liu N , Rong X , Ruan J , Huang Y : Effects of simulated microgravity and spaceflight on morphological differentiation and secondary metabolism of Streptomyces coelicolor A3(2). Appl. Microbiol. Biotechnol. 99, 4409–4422 (2015)
Jancik S , Drabek J , Radzioch D , Hajduch M : Clinical relevance of KRAS in human cancers. J. Biomed. Biotechnol. 2010, 150960 (2010)
Kokkinos MI , Wafai R , Wong MK , Newgreen DF , Thompson EW , Waltham M : Vimentin and epithelial-mesenchymal transition in human breast cancer – observations in vitro and in vivo. Cells Tissues Organs. 185, 191–203 (2007)
Kopp S , Slumstrup L , Corydon TJ , Sahana J , Aleshcheva G , Islam T , Magnusson NE , Wehland M , Bauer J , Infanger M , Grimm D : Identifications of novel mechanisms in breast cancer cells involving duct-like multicellular spheroid formation after exposure to the random positioning machine. Sci. Rep. 6, 26887 (2016)
Kraus A , Luetzenberg R , Abuagela N , Hollenberg S , Infanger M : Spheroid formation and modulation of tenocyte-specific gene expression under simulated microgravity. Muscles Ligaments Tendons J. 7, 411–417 (2018)
Kruger M , Melnik D , Kopp S , Buken C , Sahana J , Bauer J , Wehland M , Hemmersbach R , Corydon TJ , Infanger M , Grimm D : Fighting thyroid cancer with microgravity research. Int. J. Mol. Sci. 20, 2553 (2019)
Lane DP : Cancer. p53, guardian of the genome. Nature 358, 15–16 (1992)
Li J , Yen C , Liaw D , Podsypanina K , Bose S , Wang SI , Puc J , Miliaresis C , Rodgers L , McCombie R , Bigner SH , Giovanella BC , Ittmann M , Tycko B , Hibshoosh H , Wigler MH , Parsons R : PTEN, a putative protein tyrosine phosphatase gene mutated in human brain, breast, and prostate cancer. Science 275, 1943–1947 (1997)
Li J , Zhang S , Chen J , Du T , Wang Y , Wang Z : Modeled microgravity causes changes in the cytoskeleton and focal adhesions, and decreases in migration in malignant human MCF-7 cells. Protoplasma 238, 23–33 (2009)
Li N , An L , Hang H : Increased sensitivity of DNA damage response-deficient cells to stimulated microgravity-induced DNA lesions. PLoS One 10, e0125236 (2015)
Louis F , Deroanne C , Nusgens B , Vico L , Guignandon A : RhoGTPases as key players in mammalian cell adaptation to microgravity. Biomed. Res. Int. 2015, 747693 (2015)
Martinez EM , Yoshida MC , Candelario TL , Hughes-Fulford M : Spaceflight and simulated microgravity cause a significant reduction of key gene expression in early T-cell activation. Am. J. Physiol. Regul. Integr. Comp. Physiol. 308, R480–R488 (2015)
McCaffrey LM , Macara IG : Epithelial organization, cell polarity and tumorigenesis. Trends Cell Biol. 21, 727–735 (2011)
McCormick F : KRAS as a therapeutic target. Clin. Cancer Res. 21, 1797–1801 (2015)
Nan X , Tamguney TM , Collisson EA , Lin LJ , Pitt C , Galeas J , Lewis S , Gray JW , McCormick F , Chu S : Ras-GTP dimers activate the mitogen-activated protein kinase (MAPK) pathway. Proc. Natl. Acad. Sci. U. S. A. 112, 7996–8001 (2015)
Qian A , Zhang W , Li X , Weng Y , Tian Z , Di S , Yang P , Yin D , Hu L , Xu Z , Shang P : Simulated weightlessness alters biological characteristics of human breast cancer cell line MCF. Acta Astronaut. 63, 947–958 (2008)
Riboh J , Chong AK , Pham H , Longaker M , Jacobs C , Chang J : Optimization of flexor tendon tissue engineering with a cyclic strain bioreactor. J. Hand Surg. Am. 33, 1388–1396 (2008)
Sanchez-Vega F , Mina M , Armenia J , Chatila WK , Luna A , La KC , Dimitriadoy S , Liu DL , Kantheti HS , Saghafinia S , Chakravarty D , Daian F , Gao Q , Bailey MH , Liang WW , Foltz SM , Shmulevich I , Ding L , Heins Z , Ochoa A , Gross B , Gao J , Zhang H , Kundra R , Kandoth C , Bahceci I , Dervishi L , Dogrusoz U , Zhou W , Shen H , Laird PW , Way GP , Greene CS , Liang H , Xiao Y , Wang C , Iavarone A , Berger AH , Bivona TG , Lazar AJ , Hammer GD , Giordano T , Kwong LN , McArthur G , Huang C , Tward AD , Frederick MJ , McCormick F , Meyerson M , Cancer Genome Atlas Research N , Van Allen EM , Cherniack AD , Ciriello G , Sander C , Schultz N : Oncogenic signaling pathways in the cancer genome atlas. Cell 173, 321–37.e10 (2018)
Satelli A , Li S : Vimentin in cancer and its potential as a molecular target for cancer therapy. Cell. Mol. Life Sci. 68, 3033–3046 (2011)
Schmit K , Michiels C : TMEM proteins in cancer: a review. Front. Pharmacol. 9, 1345 (2018)
Shah M , Huang D , Blick T , Connor A , Reiter LA , Hardink JR , Lynch CC , Waltham M , Thompson EW : An MMP13-selective inhibitor delays primary tumor growth and the onset of tumor-associated osteolytic lesions in experimental models of breast cancer. PLoS One 7, e29615 (2012)
Sharma R , Sharma R , Khaket TP , Dutta C , Chakraborty B , Mukherjee TK : Breast cancer metastasis: putative therapeutic role of vascular cell adhesion molecule-1. Cell. Oncol. (Dordr.). 40, 199–208 (2017)
Shi ZX , Rao W , Wang H , Wang ND , Si JW , Zhao J , Li JC , Wang ZR : Modeled microgravity suppressed invasion and migration of human glioblastoma U87 cells through downregulating store-operated calcium entry. Biochem. Biophys. Res. Commun. 457, 378–384 (2015)
Takeda M , Magaki T , Okazaki T , Kawahara Y , Manabe T , Yuge L , Kurisu K : Effects of simulated microgravity on proliferation and chemosensitivity in malignant glioma cells. Neurosci. Lett. 463, 54–59 (2009)
Tan X , Xu A , Zhao T , Zhao Q , Zhang J , Fan C , Deng Y , Freywald A , Genth H , Xiang J : Simulated microgravity inhibits cell focal adhesions leading to reduced melanoma cell proliferation and metastasis via FAK/RhoA-regulated mTORC1 and AMPK pathways. Sci. Rep. 8, 3769 (2018)
Taneja P , Maglic D , Kai F , Zhu S , Kendig RD , Fry EA , Inoue K : Classical and novel prognostic markers for breast cancer and their clinical significance. Clin. Med. Insights Oncol. 4, 15–34 (2010)
Venkitaraman AR : Cancer susceptibility and the functions of BRCA1 and BRCA2. Cell 108, 171–182 (2002)
Vorselen D , Roos WH , MacKintosh FC , Wuite GJ , van Loon JJ : The role of the cytoskeleton in sensing changes in gravity by nonspecialized cells. FASEB J. 28, 536–547 (2014)
Warnke E , Pietsch J , Wehland M , Bauer J , Infanger M , Gorog M , Hemmersbach R , Braun M , Ma X , Sahana J , Grimm D : Spheroid formation of human thyroid cancer cells under simulated microgravity: a possible role of CTGF and CAV1. Cell Commun. Signal. 12, 32 (2014)
Weigelt B , Ghajar CM , Bissell MJ : The need for complex 3D culture models to unravel novel pathways and identify accurate biomarkers in breast cancer. Adv. Drug Deliv. Rev. 69–70, 42–51 (2014)
Wuest SL , Richard S , Kopp S , Grimm D , Egli M : Simulated microgravity: critical review on the use of random positioning machines for mammalian cell culture. Biomed. Res. Int. 2015, 971474 (2015)
Yin Y , Shen WH : PTEN: a new guardian of the genome. Oncogene 27, 5443–5453 (2008)
Yu XQ , De Angelis R , Luo Q , Kahn C , Houssami N , O’Connell DL : A population-based study of breast cancer prevalence in Australia: predicting the future health care needs of women living with breast cancer. BMC Cancer 14, 936 (2014)
Zhang Z , Jiang G , Yang F , Wang J : Knockdown of mutant K-ras expression by adenovirus-mediated siRNA inhibits the in vitro and in vivo growth of lung cancer cells. Cancer Biol. Ther. 5, 1481–1486 (2006)
Zheng H , Tian W , Yan H , Jiang H , Liu S , Yue L , Han F , Wei L , Chen X , Li Y : Expression of estrogen receptor α in human breast cancer cells regulates mitochondrial oxidative stress under simulated microgravity. Adv. Space Res. 49, 1432–1440 (2012)