Rujifang inhibits triple-negative breast cancer growth via the PI3K/AKT pathway.

ETHNOPHARMACOLOGICAL RELEVANCE: Rujifang (RJF) constitutes a traditional Chinese medicinal compound extensively employed in the management of triple-negative breast cancer (TNBC). However, information regarding its potential active ingredients, antitumor effects, safety, and mechanism of action remains unreported. AIM OF THE STUDY: To investigate the efficacy and safety of RJF in the context of TNBC. MATERIALS AND METHODS: We employed the ultra high-performance liquid chromatography-electrospray four-pole time-of-flight mass spectrometry technique (UPLC/Q-TOF-MS/MS) to scrutinize the chemical constituents of RJF. Subcutaneously transplanted tumor models were utilized to assess the impact of RJF on TNBC in vivo. Thirty female BLAB/c mice were randomly divided into five groups: the model group, cyclophosphamide group, and RJF high-dose, medium-dose, and low-dose groups. A total of 1 × 106 4T1 cells were subcutaneously injected into the right shoulder of mice, and they were administered treatments for a span of 28 days. We conducted evaluations on blood parameters, encompassing white blood cell count (WBC), red blood cell count (RBC), hemoglobin (HGB), platelet count (PLT), neutrophils, lymphocytes, and monocytes, as well as hepatorenal indicators including alkaline phosphatase (ALP), glutamate oxaloacetate transaminase (GOT), glutamate pyruvate transaminase (GPT), albumin, and creatinine (CRE) to gauge the safety of RJF. Ki67 and TUNEL were detected via immunohistochemistry and immunofluorescence, respectively. We prepared RJF drug-containing serum for TNBC cell lines and assessed the in vitro inhibitory effect of RJF on tumor cell growth through the CCK8 assay and cell cycle analysis. RT-PCR was employed to detect the mRNA expression of cyclin-dependent kinase and cyclin-dependent kinase inhibitors in tumor tissues, and Western blot was carried out to ascertain the expression of cyclin and pathway-related proteins. RESULTS: 100 compounds were identified in RJF, which consisted of 3 flavonoids, 24 glycosides, 18 alkaloids, 3 amino acids, 8 phenylpropanoids, 6 terpenes, 20 organic acids, and 18 other compounds. In animal experiments, both CTX and RJF exhibited substantial antitumor effects. RJF led to an increase in the number of neutrophils in peripheral blood, with no significant impact on other hematological indices. In contrast, CTX reduced red blood cell count, hemoglobin levels, and white blood cell count, while increasing platelet count. RJF exhibited no discernible influence on hepatorenal function, whereas Cyclophosphamide (CTX) decreased ALP, GOT, and GPT levels. Both CTX and RJF reduced the expression of Ki67 and heightened the occurrence of apoptosis in tumor tissue. RJF drug-containing serum hindered the viability of 4T1 and MD-MBA-231 cells in a time and concentration-dependent manner. In cell cycle experiments, RJF diminished the proportion of G2 phase cells and arrested the cell cycle at the S phase. RT-PCR analysis indicated that RJF down-regulated the mRNA expression of CDK2 and CDK4, while up-regulating that of P21 and P27 in tumor tissue. The trends in CDKs and CDKIs protein expression mirrored those of mRNA expression. Moreover, the PI3K/AKT pathway displayed downregulation in the tumor tissue of mice treated with RJF. CONCLUSION: RJF demonstrates effectiveness and safety in the context of TNBC. It exerts anti-tumor effects by arresting the cell cycle at the S phase through the PI3K-AKT pathway.

In vivo flow cytometry reveals an anti-metastatic effect of Rujifang in triple-negative breast cancer.

Breast cancer is the most common cancer, and triple-negative breast cancer (TNBC) has the highest metastasis and mortality rate among all breast cancer subtypes. Rujifang is a traditional Chinese medicine formula with many years of clinical application in breast cancer treatment. Here, we aim to investigate the effects of Rujifang on circulating tumor cell (CTC) dynamics and the tumor microenvironment in a ZsGreen/luciferase double-labeled TNBC orthotopic model. We report that the number of CTCs monitored by in vivo flow cytometry (IVFC) strongly correlates with disease progression. Rujifang treatment decreased the number of CTCs and suppressed the distant metastasis of TNBC. Moreover, immunofluorescence analysis revealed that Rujifang treatment could affect the tumor microenvironment by downregulating Kindlin-1, which has been reported to promote metastasis of TNBC. Our study provides evidence of the anti-metastatic effect of Rujifang against TNBC in an animal model using fluorescent cell lines. The results suggest the potential therapeutic value of Rujifang as an anti-metastatic drug, however, further clinical trials are needed to validate these findings in humans.

[Effects of Modified Ruji Recipe in Preventing Relapse and Metastasis of Breast Cancer Patients with Negative Hormone Receptor].

Objective To observe the effect of Ruji Recipe (RR) (treated by syndrome typing) in preventing the relapse and metastasis of invasive ductal breast cancer patients with negative hormone receptor (HR) after surgery and chemotherapy. Methods Using a prospective, cohort method, 136 pa- tients with stage I - III C HR negative invasive ductal breast cancer were equally assigned to the treat- ment group (treated by RR in syndrome typing way) and the control group (routine follow-ups). Disease free survival (DFS) , overall survival (OS) , relapse and metastasis were observed in the two groups. Re- sults All patients were followed-up for 15 to 57 months, with the median follow-up of 44 months. The median DFS and OS had not reached. The 1. 0, 1. 5, 2. -0, and 3. 0 years DFS were 94.1 % (64/68) , 86. 4 % (51/59), 81. 8% (45/55), and 72. 0% (36/50) in the treatment group. They were 77. 9% (53/68), 67.2% (45)67), 60. 6% (40)66), and 54. 5% (36/66) in the control group. Significant difference existed in 1. 0, 1. 5, and 2. 0 years DFS between the two groups (X² = 7.403, 6.426, 6.459; P =0. 012, 0.013, 0. 016). No statistical difference existed in 1. 0, 1. 5, 2. 0, and 3. 0 years OS between the two groups (P >0. 05). Among triple negative breast cancer (TNBC) patients, 1. 0, 1. 5, 2. 0, and 3. 0 years DFS were 97. 0% (32/33), 92. 9% (26/28), 92.6% (2527), and 84. 6% (22/26) in the treatment group, 81. 5% (2227), 66. 7% (1827), 61. 5% (16/26), and 57. 7% (15/26) in the control group. Of them, significant difference existed in 1. 5, 2. 0, and 3. 0 years DFS between the two groups (X² =5. 893, 7. 293, 4. 591 ; P = 0. 015, 0. 007, 0. 032). At the end of follow-ups, relapse and metastasis occurred in 15 patients, local recur- rence in 2 patients, single organ metastasis in 6 patients, and multiple organs metastasis in 7 patients of the treatment group. The relapse and metastasis occurred in 30 patients , local recurrence in 2 patients , single organ metastasis in 12 patients, and multiple organs metastasis in 16 patients of the treatment group. Conclusions RR ( by syndrome typing) could improve DFS and delay progression of invasive ductal breast cancer patients with negative HR in the first 2 years after surgery. It also had certain value for relapse and metastasis of TNBC patients within 2 years.

Treatment of middle/late stage primary hepatic carcinoma by Chinese medicine comprehensive therapy: A prospective randomized controlled study.

OBJECTIVE: To observe the efficacy of Chinese medicine comprehensive therapeutic project in treating the middle/late stage primary hepatic carcinoma (PHC). METHODS: With prospective randomized controlled design, 97 patients with PHC were assigned to the test group (49 cases) treated with Chinese medicine comprehensive therapy using Oleum fructus bruceas intervention combining oral intake of Ganji Decoction and external application of Ailitong, and the control group (48 cases) treated with chemotherapeutic agents combining iodized oil chemo-embolization and analgesics. The immediate and long-term efficacy, adverse reaction, pain-relieving initial time (PRIT) and pain-relieving sustained time (PRST) of the treatment, as well as the change in relieving patients' quality of life (QOL) were observed. RESULTS: The difference between the two groups in illness control rate was statistically insignificant (P>0.05), but the adverse reaction occurrence rate in the test group was lesser than that in the control group (P<0.05). PRIT was insignificantly different in the two groups (P>0.05), but the PRST was significantly superior in the test group than that in the control group (10.37+/-2.18 h vs 7.78+/-1.95 h, P<0.01). After treatment, the increased Karnofsky scores in the test group indicated that the patients' somatic activity, symptoms and QOL were improved significantly, which were significantly superior to those in the control group (P<0.05). The survival rate in the two groups was similar at the 3rd month after treatment, but the test group did show superiority in terms of half- and 1-year survival rate (65.9% vs 42.5% and 38.6% vs 18.1%, respectively, P<0.05). The median survival time in the test group was 8.9 months and that in the control group was 5.3 months. CONCLUSION: Chinese medicine comprehensive therapy is an effective treatment for the middle/late stage patients of PHC, and it could extend the PRST, improve the patients' QOL and long-term survival with less adverse reaction.