To investigate the antitumor activity induced by Ganoderma lucidum spore oil on different tumor cells. Human hepatoma cells(HepG2), human non-small cells lung cancer cells(A549)and human colon cancer cells(HCT116)were selected and tested. Cell viability was determined by MTT assay. Western blot analysis was performed to measure the expression of NF-κB and Caspase-3 activity in order to elucidate the mechanism of apoptotic activity caused by Ganoderma lucidum spore oil and to screen out the most sensitive cancer cell lines to Ganoderma lucidum spore oil. MTT assay demonstrated that Ganoderma lucidum spore oil had a strong inhibitory effect on the growth of three cancer cell lines. Among these cells, A549 cells were most sensitive to Ganoderma lucidum spore oil, followed by HepG2 cells and then by HCT116 cells. The results of Western blot showed that Ganoderma lucidum spore oil could promote the activation of NF-κB pathway, and that the activation of NF-κB signaling pathway in cancer cells treated by Ganoderma lucidum spore oil was stronger in A549 cells, HepG2 cells, HCT116 cells respectively. The detection of Caspase-3 activity showed that ganoderma spore oil could activate Caspase-3 dependent apoptosis pathways, which was more important in A549 cells, HepG2 cells and HCT116 cells. This study found that Ganoderma lucidum spore oil had inhibitory effects on A549, HepG2 and HCT116 cells growth and that its antitumor activity was in time-dose dependence. The mechanism may be related to the activation of NF-κB pathway and the Caspase-3 apoptotic pathway, which could accelerate apoptosis and necrosis of tumor cells. Among the three kinds of cancer cells, A549 cells was most sensitive to Ganoderma lucidum spore oil, followed by the HepG2 cells, and then by HCT116 cells.

OBJECTIVETo explore the effect of warming needling pretreatment for endometrial receptivity before frozen-thawed embryo transfer (FET).

METHODSFifty-six repeatedly embryo transfer (ET) failure patients with ultrasound showing follicular phase endometrium of C type, hysteroscopy examination presenting endometritis were randomly assigned into an observation group (25 cases) and a control group (31 cases). The patients in the observation group three months before ET were treated with antibiotics in the menstrual period, warming needle (once a day) at Zhongwan (CV 12), Tianshu (ST 25), Guanyuan (CV 4), Zhongji (CV 3), Zigong (EX-CA 1), Liangu (ST 34), Zusanli (ST 36), Shangjuxu (ST 37), Xiajuxu (ST 39) after menstruation until the ovulation stopped, and oral administration of progesterone was applied after ovulation. The patients in the control group three months before ET were treated with antibiotics in the menstrual period, and oral administration of progesterone was applied after ovulation. Continuous three menstrual periods were carried out for the both groups. The changes of endometrial thickness, type and endometrial blood flow and the outcome of FET were observed.

RESULTSEndometrial morphology and blood flow were improved after treatment in the two groups (all<0.01), with better results in the observation group (both<0.01). The embryo transplantation rate and pregnancy rate in the observation group were higher than those in the control group (both<0.01), and the early abortion rate decreased (<0.01).

CONCLUSIONSWarm needling may improve endometrial receptivity, embryo transplantation rate and pregnancy rate and decrease early abortion rate by regulating endometrial morphology and blood flow.

Insulin-like growth factor-1 receptor (IGF-1R) has been made an attractive anticancer target due to its overexpression in cancers. However, targeting it has often produced the disappointing results as the role played by cross talk with numerous downstream signalings. Here, we report a disobliging IGF-1R signaling which promotes growth of cancer through triggering the E3 ubiquitin ligase MEX3A-mediated degradation of RIG-I. The active β-arrestin-2 scaffolds this disobliging signaling to talk with MEX3A. In response to ligands, IGF-1Rβ activated the basal βarr2 into its active state by phosphorylating the interdomain domain on Tyr64 and Tyr250, opening the middle loop (Leu130‒Cys141) to the RING domain of MEX3A through the conformational changes of βarr2. The models of βarr2/IGF-1Rβ and βarr2/MEX3A could interpret the mechanism of the activated-IGF-1R in triggering degradation of RIG-I. The assay of the mutants βarr2^Y64A and βarr2^Y250A further confirmed the role of these two Tyr residues of the interlobe in mediating the talk between IGF-1Rβ and the RING domain of MEX3A. The truncated-βarr2 and the peptide ATQAIRIF, which mimicked the RING domain of MEX3A could prevent the formation of βarr2/IGF-1Rβ and βarr2/MEX3A complexes, thus blocking the IGF-1R-triggered RIG-I degradation. Degradation of RIG-I resulted in the suppression of the IFN-I-associated immune cells in the TME due to the blockade of the RIG-I-MAVS-IFN-I pathway. Poly(I:C) could reverse anti-PD-L1 insensitivity by recovery of RIG-I. In summary, we revealed a disobliging IGF-1R signaling by which IGF-1Rβ promoted cancer growth through triggering the MEX3A-mediated degradation of RIG-I.

The well-known insulin-like growth factor 1 (IGF1)/IGF-1 receptor (IGF-1R) signaling pathway is overexpressed in many tumors, and is thus an attractive target for cancer treatment. However, results have often been disappointing due to crosstalk with other signals. Here, we report that IGF-1R signaling stimulates the growth of hepatocellular carcinoma (HCC) cells through the translocation of IGF-1R into the ER to enhance sarco-endoplasmic reticulum calcium ATPase 2 (SERCA2) activity. In response to ligand binding, IGF-1Rβ is translocated into the ER by β-arrestin2 (β-arr2). Mass spectrometry analysis identified SERCA2 as a target of ER IGF-1Rβ. SERCA2 activity is heavily dependent on the increase in ER IGF-1Rβ levels. ER IGF-1Rβ phosphorylates SERCA2 on Tyr⁹⁹⁰ to enhance its activity. Mutation of SERCA2-Tyr⁹⁹⁰ disrupted the interaction of ER IGF-1Rβ with SERCA2, and therefore ER IGF-1Rβ failed to promote SERCA2 activity. The enhancement of SERCA2 activity triggered Ca²⁺_ER perturbation, leading to an increase in autophagy. Thapsigargin blocked the interaction between SERCA2 and ER IGF-1Rβ and therefore SERCA2 activity, resulting in inhibition of HCC growth. In conclusion, the translocation of IGF-1R into the ER triggers Ca²⁺_ER perturbation by enhancing SERCA2 activity through phosphorylating Tyr⁹⁹⁰ in HCC.