shPLCε Inhibits Serine/Glycine Metabolism and Proliferation of Prostate Cancer via YAP Signaling Pathway

doi:10.13523/j.cb.20191101

China Biotechnology

2019, Vol. 39

Issue (11): 1-12 DOI: 10.13523/j.cb.20191101

shPLCε Inhibits Serine/Glycine Metabolism and Proliferation of Prostate Cancer via YAP Signaling Pathway

DUAN Li-mei¹,YANG Jin-xiao¹,LIU Jia-yu²,ZHENG Yong-bo²,WU Xiao-hou²,LUO Chun-li^1,^**(

)

1 Key Laboratory of Clinical Diagnostics Founded by Ministry of Education, College of Laboratory,Chongqing Medical University, Chongqing 400016, China
2 The First Affiliated Hospital of Chongqing, Medical University, Chongqing 400016, China

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Abstract

Objective: To investigate the effects of phospholipase C epsilon on serine/glycine metabolism and cell proliferation in prostate cancer cells.Methods: Lentivirus and plasmid were transfected into LNCAP and PC3 cells. The expression of YAP, serine/glycine producing enzyme (PSAT1,PSPH,SHMT2) and proliferation-related genes (Cyclin D1,PCNA) were detected by q-PCR and Western blot. The cloning formation experiment and MTT assays were used to detect the clone formation rate and cell proliferation activity.Results: (1) Infection with LV-shPLCε significantly down-regulated the mRNA and protein levels of PLCε,YAP,serine/glycine producing enzymes (PSAT1,PSPH,SHMT2) and proliferation genes (Cyclin D1,PCNA) in prostate cancer cells LNCAP and PC3. At the same time, it inhibits the clone formation ability and proliferative activity of LNCAP and PC3 cells. (2) After adding over-expressing YAP plasmid to shPLCε group, YAP,serine/glycine producing enzymes and proliferation genes were significantly reversed. but the results of the interference with the down YAP plasmid were reversed.Conclusion: shPLCε inhibits the serine/glycine metabolism and proliferation in prostate cancer cells by down-regulating the expression of YAP.

Key words： Prostate cancer PLCε YAP Serine/Glycine metabolism Proliferation

Received: 15 April 2019 Published: 17 December 2019

ZTFLH:

Q814

Corresponding Authors: Chun-li LUO E-mail: luochunli79@126.com

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	Li-mei DUAN
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	Jia-yu LIU
	Yong-bo ZHENG
	Xiao-hou WU
	Chun-li LUO

Cite this article:

DUAN Li-mei,YANG Jin-xiao,LIU Jia-yu,ZHENG Yong-bo,WU Xiao-hou,LUO Chun-li. shPLCε Inhibits Serine/Glycine Metabolism and Proliferation of Prostate Cancer via YAP Signaling Pathway. China Biotechnology, 2019, 39(11): 1-12.

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https://manu60.magtech.com.cn/biotech/10.13523/j.cb.20191101 OR https://manu60.magtech.com.cn/biotech/Y2019/V39/I11/1

Table 1 lentivirus sequence

Table 2 The sequence of primer for q-PCR

Table 3 Plasmid vector sequence

Fig.1 PLCε mRNA and protein levels in LNCAP and PC3 cells infected with LV-shPLCε (a)mRNA level of PLCε in LNCAP infected with shPLCε by q-PCR (b)mRNA level of PLCε in PC3 infected with shPLCε by q-PCR (c)Western blot of PLCεin LNCAP and PC3 (d)Relative protein expression of PLCεin LNCAP and PC3 ^* P<0.05, ^** P<0.01, ^*** P<0.001 vs shNC group

Fig.2 Clone formation experiments of LNCAP and PC3

Fig.3 MTT experiments of LNCAP and PC3 after 72 hours (a)MTT of LNCAP (b)MTT of PC3 ^* P<0.05, ^** P<0.01, ^*** P<0.001

Fig.4 PLCε,YAP,PSAT1,PSPH,SHMT2,Cyclin D1,PCNA mRNA levels in LNCAP and PC3 cells infected with shPLCε (a) mRNA levels of LNCAP infected with shPLCε (b) mRNA levels of PC3 infected with shPLCε ^* P<0.05, ^** P<0.01, ^*** P<0.001

Fig.5 YAP,PSAT1,PSPH,SHMT2,Cyclin D1,PCNA protein levels in LNCAP and PC3 cells infected with shPLCε (a) Western blot of LNCAP infected with shPLCε (b)Relative protein level of LNCAP (c)Western blot of PC3 infected with shPLCε (d)Relative protein level of PC3 ^*P<0.05, ^** P<0.01, ^*** P<0.001

Fig.7 PLCε,YAP,PSAT1,PSPH,SHMT2,Cyclin D1,PCNA mRNA levels in LNCAP and PC3 cells infected with over-expression or knockdown of YAP plasmid (a)mRNA level of LNCAP infected with plasmids (b) mRNA level of PC3 infected with plasmids ^* P<0.05, ^** P<0.01, ^*** P<0.001

Fig.8 PLCε,YAP,PSAT1,PSPH,SHMT2,Cyclin D1,PCNA protein levels in LNCAP and PC3 cells infected with over-expression or knockdown of YAP plasmid (a)Western blot of PLCε,YAP,PSAT1,PSPH,SHMT2,Cyclin D1,PCNA in LNCAP (b)Relative protein level of LNCAP (c)Western blot of PLCε,YAP,PSAT1,PSPH,SHMT2,Cyclin D1,PCNA in PC3 (d)Relative protein level of PC3 ^* P<0.05, ^** P<0.01, ^*** P<0.001


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